Archive for the ‘PCSK9 Inhibitor Therapy’ Category

 Cholesterol Lowering Novel PCSK9 drugs: Praluent [Sanofi and Regeneron] vs Repatha [Amgen] – which drug cuts CV risks enough to make it cost-effective?

Reporter: Aviva Lev-Ari, PhD, RN


UPDATED on 4/5/2022

Early PCSK9 Inhibition in AMI Yields Plaque Regression



UPDATED on 1/15/2019

In the patent fight over PCSK9 inhibitors, the Supreme Court refused to hear Amgen’s appeal of a 2017 court decision allowing Sanofi and Regeneron to continue selling alirocumab (Praluent). Amgen still has a new patent trial starting in Delaware federal court next month, FiercePharma reports.

Amgen’s Repatha hits wall at SCOTUS but presses ahead—new price breaks included

Amgen has been trying since 2015 to protect its PCSK9 cholesterol drug Repatha by keeping Sanofi and Regeneron’s rival Praluent off the market, even going as far as to ask the U.S. Supreme Court to review an ongoing patent fight.

But that attempt fell short this week as SCOTUS refused to hear the company’s appeal of a 2017 court decision allowing Sanofi and Regeneron to continue selling its head-to-head rival.

Amgen isn’t giving up the fight, though. The company is prepping for a new patent trial starting in Delaware federal court next month. And it’s responding to long-standing criticism of the high cost of PCSK9 drugs, which hit the market in 2015 at list prices of about $14,000 a year.

Amgen had already brought the price of the biweekly version of Repatha down to $5,850 per year before discounts and rebates, and late Monday it said it would lower cost of the monthly injectable dose to that same level.


UPDATED on 11/13/2018

ODYSSEY OUTCOMES: Alirocumab Cost-effective at $6000 a Year

Marlene Busko

November 11, 2018

CHICAGO — Treatment with the proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitor alirocumab (Praluent, Sanofi/Regeneron) is cost-effective at $6319 a year when the willingness-to-pay threshold is the generally accepted $100,000 per quality-adjusted life-year (QALY), new research reports.

Deepak L. Bhatt, MD, MPH, Brigham and Women’s Hospital Heart and Vascular Center, Harvard Medical School, Boston, Massachusetts, presented these cost-effectiveness findings for alirocumab, based on data from the ODYSSEY OUTCOMES trial, here at the American Heart Association (AHA) 2018 Scientific Sessions

As previously reported, results from ODYSSEY OUTCOMES were presented at American College of Cardiology (ACC) 2018 Annual Scientific Session in March and the study was published November 7 in the New England Journal of Medicine.

Strengths of the current cost analysis include that it used actual trial data as opposed to modeling estimates, Bhatt pointed out to theheart.org | Medscape Cardiology.




Did Amgen’s Repatha cut CV risks enough to make it cost-effective? Analysts say no

Sanofi, Regeneron’s Praluent pulls off PCSK9 coup with 29% cut to death risks in most vulnerable patients
SEE our curations on PCSK9 drugs:

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ODYSSEY Outcomes trial evaluating the effects of a PCSK9 inhibitor, alirocumab, on major cardiovascular events in patients with an acute coronary syndrome to be presented at the American College of Cardiology meeting on March 10.

Reporter: Aviva Lev-Ari, PhD, RN


For PCSK9 inhibitors, the effect on major adverse cardiovascular events has always fallen short of expectations based on cholesterol lowering.

But cardiovascular risk reduction is complicated. There is more to the puzzle than cholesterol. Some drugs lower both cholesterol and prevent cardiovascular events, but some people think that the two effects are actually not that closely related.

Milton Packer MD


In a previous trial (FOURIER), another PCSK9 inhibitor had only a modest benefit on its primary endpoint, and it did not reduce cardiovascular death, although the magnitude of cholesterol lowering was striking.

In another trial (SPIRE), a third PCSK9 inhibitor, the clinical trial was terminated prematurely by Pfizer because of reduction of the effect of the drug (a humanized but not fully humanized antibody) due to development of neutralizing antibodies in some of the patients. Actually, in patients treated for more than a year who did not develop neutralizing antibodies, a beneficial effect was seen.

The ODYSSEY Outcomes trial is evaluating the effects of a PCSK9 inhibitor,alirocumab, on major cardiovascular events in patients with an acute coronary syndrome within the prior year. The drug lowers serum cholesterol dramatically, and some are hopeful that that effect will translate into an important reduction in the risk of major adverse cardiovascular events. If you believe that cholesterol reduction inevitably leads to the prevention of cardiovascular death, myocardial infarction and stroke, then you would have high expectations for the ODYSSEY trial.

ODYSSEY. The trial uses a somewhat more aggressive treatment strategy and has a longer follow-up period than its predecessors. So maybe the benefit will be large. Maybe the drug will even reduce cardiovascular death or all-cause mortality.

In order to enrich the population for cardiovascular events, the trial enrolled patients with an acute coronary syndrome within the prior year. These patients are at high risk of having a recurrence. The problem is that risk is not necessarily related to changes in cholesterol, especially the events occurring early in the trial. And in this type of trial, the analysis tends to give extra weight to early events.

Trials like ODYSSEY are often designed to stop early if the results are unbelievably impressive. The ODYSSEY trial wasn’t stopped early.

the patients entering the ODYSSEY trial are starting out with a serum LDL <100 mg/dL or even <90 mg/dL. Is cholesterol really playing an important role at that level, especially when compared with noncholesterol factors?



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FDA approval on 12/1/2017 of Amgen’s evolocumb (Repatha) a PCSK9 inhibitor for the prevention of heart attacks, strokes, and coronary revascularizations in patients with established cardiovascular disease

Reporter: Aviva Lev-Ari, PhD, RN


Evolocumab was first FDA approved in 2015 for patients with

  • familial hypercholesterolemia and
  • others who fail to achieve LDL cholesterol lowering through diet and maximally-tolerated statin therapy.

In the Repatha cardiovascular outcomes study (FOURIER), Repatha reduced the risk of

  • heart attack by 27 percent, the risk of
  • stroke by 21 percent and the risk of
  • coronary revascularization by 22 percent.2


U.S. Repatha Indication

Repatha is a PCSK9 (proprotein convertase subtilisin kexin type 9) inhibitor antibody indicated:

  • to reduce the risk of myocardial infarction, stroke, and coronary revascularization in adults with established cardiovascular disease.
  • as an adjunct to diet, alone or in combination with other lipid-lowering therapies (e.g., statins, ezetimibe), for treatment of adults with primary hyperlipidemia (including heterozygous familial hypercholesterolemia [HeFH]) to reduce low-density lipoprotein cholesterol (LDL-C).
  • as an adjunct to diet and other LDL‑lowering therapies (e.g., statins, ezetimibe, LDL apheresis) in patients with homozygous familial hypercholesterolemia (HoFH) who require additional lowering of LDL‑C.

The safety and effectiveness of Repatha have not been established in pediatric patients with HoFH who are younger than 13 years old.

The safety and effectiveness of Repatha have not been established in pediatric patients with primary hyperlipidemia or HeFH.

Eligible patients with high cholesterol (LDL-C ≥70 mg/dL or non-high-density lipoprotein cholesterol [non-HDL-C] ≥100 mg/dL) and established cardiovascular disease at more than 1,300 study locations around the world were randomized to receive Repatha subcutaneous 140 mg every two weeks or 420 mg monthly plus high- or moderate-intensity effective statin dose; or placebo subcutaneous every two weeks or monthly plus high- to moderate-intensity statin dose. Statin therapy was defined in the protocol as at least atorvastatin 20 mg or equivalent daily with a recommendation for at least atorvastatin 40 mg or equivalent daily where approved. The study was event driven and continued until at least 1,630 patients experienced a key secondary endpoint.

About Repatha® (evolocumab)
Repatha® (evolocumab) is a human monoclonal antibody that inhibits proprotein convertase subtilisin/kexin type 9 (PCSK9). Repatha binds to PCSK9 and inhibits circulating PCSK9 from binding to the low-density lipoprotein (LDL) receptor (LDLR), preventing PCSK9-mediated LDLR degradation and permitting LDLR to recycle back to the liver cell surface. By inhibiting the binding of PCSK9 to LDLR, Repatha increases the number of LDLRs available to clear LDL from the blood, thereby lowering LDL-C levels.1

About Amgen in the Cardiovascular Therapeutic Area
Building on more than three decades of experience in developing biotechnology medicines for patients with serious illnesses, Amgen is dedicated to addressing important scientific questions to advance care and improve the lives of patients with cardiovascular disease, the leading cause of morbidity and mortality worldwide.8 Amgen’s research into cardiovascular disease, and potential treatment options, is part of a growing competency at Amgen that utilizes human genetics to identify and validate certain drug targets. Through its own research and development efforts, as well as partnerships, Amgen is building a robust cardiovascular portfolio consisting of several approved and investigational molecules in an effort to address a number of today’s important unmet patient needs, such as high cholesterol and heart failure.

Homozygous Familial Hypercholesterolemia (HoFH): In 49 patients with homozygous familial hypercholesterolemia studied in a 12-week, double-blind, randomized, placebo-controlled trial, 33 patients received 420 mg of Repatha subcutaneously once monthly. The adverse reactions that occurred in at least 2 (6.1 percent) Repatha-treated patients and more frequently than in placebo-treated patients, included upper respiratory tract infection (9.1 percent versus 6.3 percent), influenza (9.1 percent versus 0 percent), gastroenteritis (6.1 percent versus 0 percent), and nasopharyngitis (6.1 percent versus 0 percent).

Immunogenicity: Repatha is a human monoclonal antibody. As with all therapeutic proteins, there is a potential for immunogenicity with Repatha.

Please contact Amgen Medinfo at 800-77-AMGEN (800-772-6436) or 844-REPATHA (844-737-2842) regarding Repatha® availability or find more information, including full Prescribing Information, at www.amgen.com and www.Repatha.com.


  1. Repatha® U.S. Prescribing Information. Amgen.
  2. Sabatine MS, Giugliano RP, Keech AC, et al, for the FOURIER Steering Committee and Investigators. N Engl J Med. Evolocumab and Clinical Outcomes in Patients with Cardiovascular Disease. 2017;376:1713-22.
  3. Cannon CP, et al. N Engl J Med. 2004;350:1495-1504.
  4. LaRosa JC, et al. N Engl J Med. 2005;352:1425-1435.
  5. Pederson TR, et al. JAMA. 2005;294:2437-2445.
  6. Search Collaborative Group Lancet 2010;376:1658–69.
  7. Cannon CP, et al. N Engl J Med. 2015;372:2387-2397.
  8. World Health Organization. Cardiovascular diseases (CVDs) fact sheet. http://www.who.int/mediacentre/factsheets/fs317/en/. Accessed October 30, 2017.



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Trends in HealthCare Economics: Average Out-of-Pocket Costs, non-Generics and Value-Based Pricing, Amgen’s Repatha and AstraZeneca’s Access to Healthcare Policies

Reporter: Aviva Lev-Ari, PhD, RN


1.   AstraZeneca’s access to healthcare strategy is made up of three elements:

  • Provide high-quality, effective and appropriate medicines to those who need them. Improve affordability, particularly among the growing middle class in Emerging Markets.
  • Bring down healthcare barriers, particularly in developing countries. Our strategy helps us to address affordability and other healthcare barriers, while ensuring we continue to provide high-quality medicines to those who need them.
  • Key Target exceeded Full target achieved Ongoing progress Target not achieved, some progress AstraZeneca has extensively expanded and updated their access strategy identifying those areas where they are best placed to provide support and are now well positioned for future progress.”
  • Access to Medicine Index Access to healthcare Goals Target progress Progress highlights Expand sustainable patient access to our medicines to reach 3 million patients by 2016 4.49 million patients in Emerging Markets served by patient access programmes
  • Young Health Programme After exceeding initial goal to reach 1 million people through the Young Health Programme by 2015, aim to renew in five markets and expand into three markets by 2018 Renewed in Canada, Germany, China and India and expanded into Kenya
  • Total reach in 2016 of 166,000 and 1.6 million youth since 2010
  • Proposals for expansion are in development for Brazil and Australia and for renewal in Portugal
  • Healthy Heart Africa Reach 10 million hypertensive patients across Sub-Saharan Africa by 2025 Since 2014, we have conducted over 2.7 million screenings and started treatment for over 100,000 hypertensive patients



Click to access Access%20to%20healthcare.pdf

2.   Co-Development and Commercialization by Territory

AstraZeneca has paid $45 million and committed to up to $2.1 billion in milestones to team with Pieris Pharmaceuticals. The agreement sets Pieris up to move respiratory candidate PRS-060 into the clinic on AstraZeneca’s dime and pull in milestones as it and other pipeline prospects advance.

Tiny Pieris is due to receive the first, $12.5 million milestone when it moves moderate to severe asthma candidate PRS-060 into phase 1. AstraZeneca will fund clinical development of the interleukin-4 receptor alpha-targeting protein. If the asset reaches phase 2a, Pieris has the option to codevelop and commercialize it in the U.S., bumping up the royalties or gross margin share it receives in the process.

Pieris has a similar codevelopment option on other assets covered by the agreement. The biotech will develop four other proteins against undisclosed respiratory targets. If Pieris wants, it can sign up to codevelop and commercialize two of these programs in the U.S. Milestones and commercial payments across the deal could ultimately total $2.1 billion.



3.  Prescriptions Dispensed at Zero Patient Out-of-Pocket Cost Reached Thirty Percent in 2016

29.9% of prescriptions have been dispensed at zero patient out-of-pocket cost, including brands and generics, up 1.5% since 2015, all due to increased use of zero cost generics.
The total share of prescriptions where patients paid some amount less than $50 declined by 1.3% to 67.8% in 2016.
The proportion of claims with patient cost exposure greater than $50 increased also declined slightly from 2.5% to 2.3% in 2016.

Since 2013, Average Out-of-Pocket Costs for All Brand and Generic Prescriptions has Decreased by $1.19

Average patient out of pocket costs declined from $9.66 in 2013 to $8.47 in 2016, with 2016 brand costs declining to $28.31 from $32.36 in 2013 and generics dipping to $5.54 from a high of $6.05 in 2013.
The list prices of brands continue to be far higher than the average paid by patients, as few patients are exposed to those costs in their insurance plans.
The average list price for brands averaged 12 times higher than the average out of pocket cost for patients in 2016 compared to 3 times higher for generics.


For Immediate Release Contact: Joan Fallon

May 2, 2017 joan_fallon@harvardpilgrim.org


4.   Harvard Pilgrim Signs Second Groundbreaking Contract with Amgen For Repatha

HPHC and its members will receive full refund if a cardiac event occurs while on the drug

(WELLESLEY, MA) – Harvard Pilgrim Health Care has entered into a first-of-its-kind contract with Amgen for its LDL cholesterol lowering drug, Repatha, that guarantees the health plan and its members will receive a full refund of their costs for the drug if a member is hospitalized for a myocardial infarction or stroke after taking Repatha for six months or more and maintaining an appropriate level of compliance on the drug.

Repatha is one of a new class of biotechnology medicines known as PCSK9 inhibitors that have demonstrated a promising new approach for treating elevated LDL cholesterol in patients whose levels are not able to be controlled by current treatment options. The medication is designed to target a protein that prevents the body from removing artery-blocking LDL cholesterol from the bloodstream. Repatha works differently than statin drugs that prevent the liver from making cholesterol.

Given by injection every two or four weeks, Repatha is intended for patients who have an inherited disorder resulting in high levels of LDL cholesterol or have high-risk atherosclerotic cardiovascular disease conditions, such as heart attack or stroke, that have been resistant to treatment.

“Repatha has been shown to have a significant outcome on reducing cardiovascular morbidity for high risk individuals with elevated LDL cholesterol,” said Harvard Pilgrim Chief Medical Officer Michael Sherman. However, there have been concerns raised about the cost of this new drug relative to existing statin treatments. We hope to negotiate more contracts of this type, in which a pharmaceutical company truly has ‘skin in the game’ going forward. This agreement is the first we have signed in which there is a full refund of all costs related to the medication if the patient experiences a heart attack or stroke while taking it.”

“Cardiovascular disease is the largest public health concern in the world and for high-risk patients who have already had a cardiovascular event or whose genetics puts them at risk, it is important that these patients have access to an effective treatment shown to lower their

elevated LDL cholesterol in addition to their current lipid lowering regimen,” said Joshua J. Ofman, M.D., MSHS, senior vice president of Global Value, Access & Policy. “Amgen’s agreement with Harvard Pilgrim demonstrates our commitment to seeking innovative approaches that help break down the barriers of access to Repatha.”

This is the second patient-focused outcomes contract Harvard Pilgrim has negotiated with Amgen for Repatha. In the fall of 2015, the health plan signed an outcomes guarantee through which Amgen provided HPHC with an enhanced discount if the reduction in LDL levels for Harvard Pilgrim members is less than what was observed during Repatha’s clinical trials. In addition, the agreement provides for additional discounts if the utilization of the drug exceeds certain levels. This enables those patients who can most benefit from the drug to receive it while continuing to encourage utilization of lower cost statins for the majority of patients.


From: “Fallon, Joan” <joan_fallon@harvardpilgrim.org>

Date: Tuesday, May 2, 2017 at 1:09 PM

Subject: press release from Harvard Pilgrim Health Care

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Highlights – LIVE Day 2: World Medical Innovation Forum – CARDIOVASCULAR • MAY 1-3, 2017  BOSTON, MA • UNITED STATES


Leaders in Pharmaceutical Business Intelligence (LPBI) Group will cover the event in


Aviva Lev-Ari, PhD, RN will be streaming live from the floor of the Westin Hotel in Boston on May 1-3, 2017




Forthcoming SEVEN e-Books in 2017 AND Eight e-Books on Amazon.com


Biggest Voices in Cardiovascular Care

2017 World Medical Innovation Forum: Cardiovascular, May 1-3, 2017, Partners HealthCare, Boston, at the Westin Hotel, Boston


Tuesday, May 2, 2017

7:00 am – 8:00 am
Lilly Foyer
7:00 am – 7:45 am
Pfizer Ballroom
FOCUS SESSION: Japan Today: Advancing Cardiometabolic Therapies

Discussion on unique aspects of cardiometabolic market in Japan, its projected trend over the next 5 years and explore transformative models of open innovation to accelerate development of new therapeutic options.

  • Yoshiro Miwa Associate Chair and Founding Director, Center for Interdisciplinary Cardiovascular Sciences, Brigham and Women’s Hospital
  • Head of Pharmaceuticals, Americas Region, Bayer
  • Director, Health & Welfare Department, JETRO New York
  • President, Japan Agency for Medical Research and Development
  • President, Head of Global Business Development, Mitsubishi Tanabe Pharma Holdings America, Inc.
  1. Complications of Toxin absorption and metabolic disease
  2. Collaboration with Academia: @ representatives are on two West Coast and 2 on the East Coast
  3. CVD and HTN related to aging is on the rise National Initiative to encourage change in Life Style
7:50 am – 8:00 am
Boston Scientific Ballroom
Opening Remarks
  • Chief Innovation Officer, Partners HealthCare
8:00 am – 8:50 am
Boston Scientific Ballroom
Pricing to Enable Affordability and Innovation

Balancing acceptable answers to high and escalating drug prices in the United States while making strides in medical innovation. Leaders in innovation, policy, care delivery, academia, and insurance discuss potential collaborative solutions.


Moderator: Peter Slavin, MD
  • President, Massachusetts General Hospital
  1. American Consumer of Healthcare pays more and it is not justifiable
  2. Pay for Value, pay for Outcomes
  • Physician-in-Chief, Department of Medicine, Massachusetts General Hospital
  • Jackson Professor of Clinical Medicine, Harvard Medical School
  1. Challenges understand PCP services and SPacialty medicine
  2. Adding fluids or taking it away is the majority of the decisions
  3. In cancer treatment 40% of prescriptions are not filled due to out of pocket cost increase
  4. In drugs Innovation are more expensive not less expensive
  5. Economists: Physicians are irrational
  6. Patient engagement, own health in their hands for compliance with treatment
  7. Assist MDs with the right data for their decision on what drugs to use
  8. Two key ways : Complications of Drugs, drive drug cost – Personalized medicine – improve outcomes on an Individual Patient basis
  9. How important is the question, affordable drugs is more important than anything in the delivery of care
  • CEO, Boehringer Ingelheim USA
  1. Many stakeholders are involved
  2. Pricing of Pharmaceutical in last 10 years, “List price” and the “Net Price” collected by Pharma has widen,
  3. high deductable plans are prevalent 40%-50% – out of pocket cost increased
  4. backlog of generic drugs – it takes 36 month to approve vs 12 month of non-generic
  5. Value-based pay, drug is only one enabler in MDs tool kit
  6. Out of pocket cost: Exposure is largest on the drug-side, that is preventive to avoid hospitalization
  7. Unfair pricing leading to not be active in certain markets, Price control outside the US, take position on Importation, not disrable to import drugs into the US, we do not wish drug shortage around the world, Canada is a Small market US is a huge market
  8. FDA on Oncology drug-device, potential exists for existing drugs
  9. Continue to do Clinical Trials in the US, claims orientations exacerbation, describe the benefit
  • EVP, Medical Devices, Abbott
  1. Who will pay and why?
  2. How we challenge development team to bring down cost of technology and plans showing cost savings in 12 month not few years down the road
  3. Selection of areas: ORTHO – hip replacement and Pain management
  4. Establish Global Pricing Models in USD, Premium, fair Price, desperative Prices is not good for the system
  • Director of Innovation, Cardiovascular Division, Senior Investigator, TIMI Study Group, Brigham and Women’s Hospital
  • Associate Professor of Medicine Harvard Medical School
  1. Cardiometabolic diseases  – drugs available to avoid events down the road
  2. new drugs at $20,000 cost per year vs Generic drugs – Economic responsibility in the Lipids area is long term
  3. MI many types bundling cost is more difficult that in Ortho
  4. Chronic disease, therapeutics, diagnostics, How to reduce cost? – Best utilization of drugs
  5. Durable response to drug, not enough data in hands of MDs
  6. Randomize Placibo and Randomize the drug, Placibo – requires better engagement
  7. After MI – 6 MEDS, compliance
  9. develop platform to test simple questions, in Cardiology
8:50 am – 9:40 am
Boston Scientific Ballroom
CLINICAL HIGHLIGHT: Emerging Devices for Complex Structural Heart Disease

Evolution of mitral disease management, current practice and impact of new technologies on both repair and replacement, implications of a heterogeneous patient population, triage, timing of intervention.

Moderator: Jason Mills
  • Managing Director, Head of US Healthcare Research, Canaccord Genuity
  1. What patient to target
  2. Heterogenous population – definitive data, how it is achieved
  • Divisional VP and General Manager, Structural Heart, Abbott
  1. Homogenous or heterogenous
  2. Standard of Care- restore normal function, patient outcomes more fragile as disease progresses
  3. Paradigm, measurable reduction of regurgitation
  4. Design of Clinical Trials: MR treatment around the World,
  5. MitralClip reduce MR reduction is not resolution
  6. 1000 publication on MitralClip – data gather indicate improvement in life quality
  7. TEE alone for use of MitralClip is nor enough, need to see to do the procedure
  • EVP, Global CMO, Boston Scientific
  1. State of the Art, Mitral regurgitation and degenerative Mitral valve: mechanism and elements responsible for regurgitation, repair of Annuals vs replacement of the valve.
  2. Options at different stage of the disease
  3. Functional Mitral Regurgitation: care pathways, compounding effects, two little too late
  4. AF can cause Valve dilatation and regurgitation
  5. Treatment, patient less symptomatic
  6. HTN cause of LV systolic disfunction – treated first – improve the Mitral regurgitation
  7. Mechanism under pinning in the decision process, CLinical Trials – Device may not work for all patients in the Study
  8. leaflet condition dealt in repair strategy vs device selection
  9. Having devices focus the clinical pathways for therapeutic options, TAILORING OF DEVICES TO SPECIFIC STRUCTURAL CONDITION OF THE HEART
  • Watkins Family Distinguished Chair in Cardiology, Brigham and Women’s Hospital
  • Professor of Medicine, Harvard Medical School
  1. Structural Heart disease: Hole in Heart,
  2. 5 1/2 years approved for TARV
  3. TAVR will exceed Open Heart Surgery next year
  4. Mitral valve growth is in  >75 y-o more cases than Aorta
  5. Aorta Valve – seen on Echogram stenosis seen
  6. Mitral Valve – concert of several elements, very complicated, Coordination among: Device, FDA, CMS, MDs, Hospitals
  7. DO  doctors wait to long to intervene: Moderate to severe: Foundational approach Ventricular dysfunction, late stage not continue to progress.
  8. MI
  9. drivers of cords,
  10. identify Patient – can be improved by PCPs, NPs, PAs, assess severity, Center for evaluate consensus on timing the intervention
  11. relay on the Cardiologist in the Community – context: Not all MR needs surgery
  12. Functional Mitral Regurgitation, poor LV function, the valve intervention will not improve longevity may improve quality of life for two years
  13. 20% survival after MI in LV dysfunction post MI then Mitral valve intervention will not improve longevity
  14. For older pation with Functional MR and moderate LV dysfunction – trial design on utility of the intervention.
  15. More patients will be included for treatment as recognition of the disease matures
  16. WHO should do that procedue : Interventional Cardiologist or Cardiac SUrgeons: HARD procedure NOT fixing Coronary artery
  17. Set up regional centers of Care links to maintain quality and PATIENT MUST DO BETTER
  18. 200 centers in the US do MitralClip procedure
  19. Procedure expensive BEYOND THE DEVICE cost


  • CVP, Advanced Technology, CSO, Edwards Lifesciences
  1. 60,000 procedures in the US vs. 2.4 million patients with the MR condition
  2. Percutanious is an opportunity not to damage the heart, challenge, how to attach  to the heart and how much regorgitation to get clinical benefit, optimal benefit to patient: Multiple products are in development
  3. Aortic stenosis: we learned which patient will benefit, clinical studies, cost effective, two companies validated the approach
  4. Mitral Valve is in early stage Trans catheter is the direction
  5. PATIENT ACCESS – who will benefit
  6. devices will Improve Patient conditions
  • SVP and President, Coronary & Structural Heart, Medtronic
  1. MR at medtronic: degenerative disease, repair the valve, average surgeon does 6 procedures a year,
  2. Toolbar approach, how to do it safely no complication repeatable to know the reduction level
  3. population exists to do the development early in the stage of MR
9:40 am – 10:10 am
Boston Scientific Ballroom
1:1 Fireside Chat: John Lechleiter, PhD, Chairman, Eli Lilly
Moderator: Susan Dentzer
  • CEO, Network for Excellence in Health Innovation
  • Chairman, Eli Lilly and Company
  1. Two approaches to Beta Ameloid – fail to meet Endpoint: Mild patient Solismad 24% improvement vs placibo
  2. Dementia not AD – mild to moderate patients, only.
  3. Move faster is desiable, turnaround time need be faster
  4. Would do over again, tap best minds in the World,
10:10 am – 10:25 am
Lilly Foyer
10:25 am – 11:15 am
Boston Scientific Ballroom
Personal Monitoring for Disease Management

Considering the evolving trends in viability and utilization and the opportunities wearables may present for real-world clinical decision making.


Moderator: Joe Kvedar, MD
  • VP, Connected Health, Partners HealthCare
  • Associate Professor of Dermatology, Harvard Medical School
  1. Evidence on monitoring Patients while @Home, pros and cons
  2. 2016, review evidence, recommendation for monitoring Patients while @Home
  3. Continuing care and continuing data collection
  4. Hospital administrator need a path to have more patients coming to the hospital
  5. Implement technology for quality care, access and lower cost
  • CIO, VP, Brigham and Women’s Hospital
  • Course Co-Director, Harvard Medical School
  1. CHF, HF, – recognize that Technology alone is not enough
  2. People and Technology intervention targeting
  3. Academic medical centers – monitoring Patients while @Home is a mechanism to deliver care
  • COO, Siemens Healthineers
  1. Outcome-based evidence – innovation exited 15 years ago
  2. at Present time the market is accepting
  3. Medical Systems do not have enough capacity – shortage of MDs
  4. Monitoring Patients while @Home is to free time of MDs in the Office
  • CEO, Zoll Medical
  1. Outcome-based research on a wearable cardio-devibrilator, Arrhythmic death protection
  2. Policy: talk about reimbursement
  3. Patient data collected, histories of ECG before cardiac arrest
  4. what diagnostics to be used with this data: do not drive, do not be alone at home
  • CSO, One Brave Idea, Brigham and Women’s Hospital
  • Associate Dean for Executive Education, Harvard Medical School
  1. Evidence and publishing results, MDs and Patient’s perspective on Autonomy vs monitoring Patients while @Home
  2. DIgital Health Comapny vs Academic Study on monitoring Patients while @Home – Wearable Patch surpass wearing a holter
  3. External wearable now acceptable and clinical evidence will convince all stakeholders
  4. Realization by physicians that monitoring Patients while @Home is a TIME SAVER in their practice will endore the technology at a rapid pace
  5. Published studies: Sharing genetic information with Academic Centers: Verily, AstraZeneca and AHA partnership
  6. Information in the Periphery but adopted in the center of Unifies healthcare eco system not in Silos anymore
11:15 am – 11:45 am
Boston Scientific Ballroom
1:1 Fireside Chat: Robert Bradway, CEO, Amgen
Moderator: Scott Sperling
  • Co-President, Thomas H. Lee Partners
  1. Amgen –>>> Biotech to Pharma
  • CEO, Amgen
  1. Six areas: CVD, Cancer, Inflammation,
  2. CVD opportunities: Science and commercial – Heart disease, tools of Human genetics for drug development in CVD: REPATA a molecule targeting PCSK9 – variant on gene associated with LDL Pathways – genetic clue
  3. Innovation in Human genetics new sequencing technologies allowed to see disease in Human populations, disease and pathways
  4. Aging associated with risks of CVD, How we pay for innovative therapies?
  5. Benefit from innovation – 800,000 in US have a stroke every year $60 Billion treatment for patient of CVD
  6. Value of innovation at a price that allows access and lowering cost of care
  7. Cardiologist prescribed the medication for himself it took 6 month for insurance to approve
  8. Utilization management – move to innovative technologies if current therapies do not work
  9. Pay for benefit and for outcome, no pay if med does not do what it was supposed to do – refund patients
  10. focus on right patient get access. if LDL is so high – the therapy is there – the payers, enable access
  11. Access challenge: Discount, Rebates, Co-pay assistance to access therapy as REPATA at $5 a day value is high,
  12. A single payer is the Government in other countries
  13. Future at Amgen: Potential for Innovation to improve Medicine, paying for innovation needs to be strainten
  14. Coming drug is Pharmcogenetics for atherosclerosis
12:00 pm
GE Ballroom
12:15 pm – 12:30 pm
GE Ballroom
Austen-Braunwald Award

Awarded to one BWH and one MGH First Look participant who embodies the innovative, entrepreneurial, and visionary spirit of cardiovascular legends W. Gerald Austen, MD and Eugene Braunwald, MD. Granted based on select criteria, including overall presentation quality, innovativeness, commercial potential, caliber of disruption, and market need.

  • Ben Olenchock, BWH
  • Steven Lubitz, MGH
12:30 pm – 1:00 pm
GE Ballroom
1:1 Fireside Chat: Frans van Houten, CEO, Philips
Moderator: Gregg Meyer, MD
  • CCO, Partners HealthCare
  1. Why Healthcare?
  2. How your approach to innovation enable to move fast?
  3. Develop technologies that are more affordable
  4. Data, Insight, How to get insight from data about a deterioration
  • CEO, Philips
  1. A 125 year company, shade lighting business to focus of Healthcare, global challenge a goal in Humanity for solution, services, products
  2. R&D diagnostics, Informatics to integrate data
  3. Africa and India – emerging markets with infant mortality high — develop a clinic as franchise for every price point
  4. shift from Products to Cloud-based solutions – Prevention, Diagnostics, @Home care: Neuro, Cancer, CVD
  5. Academic Institutions: Karlinska in Sweden – Stroke solution in partnership with Philips
  6. Affordability, maximum of the technology, partnership with Industry consultants, does not work everywhere, took in house the Services part and developed algorithms to assist MDs in interpretation of radiological data
  7. Patient monitoring 24×7 in ICUs,
  8. eICU – measure evolution to forcast 6 hours in advance a deterioration – highest performance, reduction 40% of death by insight from data
  9. Complex diseases created enormous data,
  10. Measuring progression of AM – AI algorithms for a digital platform
  11. Data integration, oncology patients: Genomics, Pathology, Clinical Data Scientist,
  12. R&D will be co-creation with clinical validation and publication for Market adoption
  13. Head of Radiology across several Hospitals – Better Outcomes Operations improvement due to technology
  14. Rural Africa market connected to a Hospital in a city — working on that teleconference
  15. UAE – crowdsource for nearest AED – locate incidence like UBER for CVD
  16. AI in Pathology – genomics and patient targeting – Lab in Cambridge, Big Data
1:00 pm – 1:10 pm
1:10 pm – 2:00 pm
Boston Scientific Ballroom
Global Clinical Trials: Next Generation Design and Scalability

Cardiovascular trials currently account for 10 percent of all clinical trial participants. Discussion on design and implementation of clinical studies globally, considering strategies for patient access, regulatory implications, cost containment and management of relationships with global service providers.


  • Chairman, TIMI Study Group, Lewis Dexter, MD Distinguished Chair in Cardiovascular Medicine, Brigham and Women’s Hospital
  1. Bar is very high, events went down, that necessitates very large studies 30,000 patients, 12 sites around the globe, acquisition of data
  2. Outcomes, where is Pharma in development of a compound
  3. Other indications that LIPIDS
  4. surrogate outcomes
  5. Diabetes: requirements mutual effect on CVD, benefit of DM drugs for CVD OutcomesGathering data on approved drugs, looking at different doses
  6. A model for dosing compounds per each patient
  7. Benefit or harm different in the US and in other sites
  8. genetics, how it inform in drug development, validation to pan out
  9. Real World evidence in 21 Affordable Act – randomize
  10. multivariable adjustment on how many variable affect the Power of the study
  • VP, Cardiovascular & Metabolic Disease Head, Global Medicines Development, AstraZeneca
  1. Patient population,
  2. new medicine vs existing registries
  3. Real World evidence include observational dat after the drug is on the market
  4. Randomization – identify population, register, randomize, collect
  • SVP, Global Clinical Research, Therapeutic Area Head, Cardiometabolic & Womens Health, MRL Lead, China R&D, Merck
  1. data from medical record
  2. investigational drugs more difficult
  • VP Cardiovascular Medicine, Covance
  1. Productive sites, approaches: Russia, Russia-Georgia, dishonest recruitment, combine resources to find sites
  2. 50% of blood analysis at sites that do not recruit at all
  3. Internal vs External validity: Early on in drug development, protocol deviation, drug MOA, variability off set by study power
  4. Patient reporting outcomes
  • Director, Division of Cardiovascular and Renal Products, Food and Drug Administration
  1. Surrogates: likely outcome endpoint for decision,
  2. CVD is difficult, graveyard of program that failed, inotropic drugs were stopped
  3. Global trial, most expensive to do in the US – in the US the care given need be comparable to the care in the US
  4. Requirement that the results will be relevant to type of care in the US
  5. Preserve randomization is key
  • VP, Cardiovascular Medicine,  Global Development, Amgen
  1. What is the hypothesis for testing, population match, understand the molecule for the modality
  2. Decode, benefits and risks – phynotype
  3. aggregation of data with investigational therapies,
  4. AI will become part of Clinical Trial
2:00 pm – 2:50 pm
Boston Scientific Ballroom
Precision Cardiovascular Medicine: What is Different This Time

Explore how precision medicine is changing the face of cardiovascular medicine specifically. The session will examine the impact of combined phenotypic and genotypic characterization on optimizing response to therapeutics, trial design, improving outcomes, and redefining reimbursement.

  • EVP, R&D, Amgen
  1. Outcomes for RAPATA – a pharmacogenomic drug
  2. Precision medicine in CVD – optimistic
  3. CVD – phynotype more determinative then genotyping vs Oncology, complex traits
  4. Bippharma moves away from big public health diseases, trials are expensive, FDA harch requirements
  5. Investors:to Biotech – works on Oncology and on Orphan drugs
  6. Methodology for targeting by using genetics are more precise
  7. In Phase III a drug where biology is very well understood
  • VP, Head Translational Medicine Merck
  1. CVD in Merck – rearrange resources in South SF on
  2. Arrhythmia: Mutation if down played causes Arrhythmia if Overexpressed causes Arrhythmia – caution in terapeutics tatgets – gene indication not to develop therapy
  3. Diastolic HF – make a drug, pick up one signaling cascade and show efficacy not in all pathways
  4. Populations that are resilient in diseases as HF
  • Assistant in Medicine, Massachusetts General Hospital
  • Assistant Professor, Harvard Medical School
  1. AF model in Translational medicine, metabolites
  2. moderately optimistic
  3. molecular phynotyping
  • Director, Cardiovascular Genetics Center, Brigham and Women’s Hospital
  • Thomas W. Smith Professor of Medicine and Genetics, Harvard Medical School
  1. Genetic in CVD – Cardiomyopathy and genetics
  2. target molecule for therapy of genetic
  3. gene mutation variants are different the genes are the same
  4. LDL receptor led to development of Statins
  5. PCSK9 was developed from genetic observation on familial
  6. protein profiles very important
  7. Genotype more informative than phynotypes
  8. Genetic tools to direct drug discovery

Kevin Hrusovsky, Quanterix

  • Biomarkers at the bedside
  • Protein of inflammation in DM – phynotype, genotype – stratify population of patients for targeting therapeutics
  • 6 inhibitions, role of protein, multiple cytokines involved
  • Head injury – diagnostics must be very quick
  • Insurance will require prevention emphasis
  • Early diagnosis is facilitated by genokmics


2:50 pm – 3:40 pm
Boston Scientific Ballroom
CV Investing in the Next Decade

View on investing landscape, opportunities in the CV/metabolic marketplace, the drugs, devices and diagnostics currently in pipelines and notable positive trends.

Moderator: Meg Tirrell
  • Reporter, CNBC
  1. M&A landscape
  • Managing Director, Healthcare, GE Ventures
  1. Advanced diagnostics
  2. value-based care
  3. no investment in drugs
  4. Insurance are into the Game of Data Analytics – fast adoption to become standard of care
  5. Reimbursement:  Tech investors and Healthcare investors with having in mind FDA approval process
  6. Mobile health cool: eye disease, DM, skin care ECG, few specialists in China, mobile tools
  7. Interoperability in Digital Health
  • Partner, Atlas Venture
  1. Only investment in drug discovery
  2. Segment genotypes – pure innovations as differentiators
  3. Patient Analytics, Physician-Patient SW development applications – scale broad audience – value add
  4. Focus on Medical Professions tool development for this sector
  5. Learning curve for novel productivity tools Cardiac MR – Imaging Analytics – Precision medicine not in drugs but in imaging
  6. M&A – activity 4 years time horizon, new biology new modality – risk is higher
  7. First in Class
  8. Translational Research and Drug discovery are two different beasts, doing drug development inside a basic research organization
  9. Coolest technology: CRISPR – one injection reduction in a genetic disease
  • Managing Director, US Medical Technology, Equity Research, Bank of America Merrill Lynch
  1. Medtech, CVD is exciting , i.e., Valve area, ICD, Stents, Stroke, AF,
  2. Medical devices – exciting
  3. No clear leader in Mitral Valve repair and Replacement by 2019 – approved products in Europe: Abbott, Medtronic (12), Edwards
  4. Value in the market exists for investors
  5. coolest technology: Stroke – stents in the barin
  • VP, Venture, Partners HealthCare
  1. 165million fund: Drugs, devices, diagnostics – ONLY from Partner COMMUNITY developed IP
  2. Orphan CVD driven by genomics
  3. Stratify the patients to show effects
  4. Exit for medical devices is longer than drugs with innovative business models
  5. Wearables are medical devices
  6. Data will be huge and valuable
  7. Skill set needed for Drug discovery and Academic science — DOES work well in one place
  8. Editas – Academic Center: Innovations everywhere
  • Managing General Partner, Frazier Healthcare Partners
  1. Drug development investment in early stage and in late stage
  2. Focus opportunities
  3. 3 to 5 years time horizon
  4. $50 – $60million investment range
  5. FDA – is central to HC investment
  6. FDA – changed regulation to enable antibiotics development
  7. FDA in Oncology – risk reward equation – FDA played great role in drug development
  8. Leukemia, non-Hodkin Limphoma
3:40 pm – 4:30 pm
Boston Scientific Ballroom
CLINICAL HIGHLIGHT: Optimizing Care for the 51%: New Market Opportunities

Introduction: Cathy Minehan, Chair, MGH Corporation

Address implications of gender as a key biological factor for personalized medicine. Stroke is likely to be the first cardiovascular event, tied to AF and secondarily to hypertension. Opportunities for medication utilization and optimization in context of, manifestation of disease and understanding the biology, complications, strategies to collect relevant clinical evidence, and treatment response.

 Nancy Brown,
  • CEO, AHA
  1. Biology or bias
  • Director, Center for Arrhythmia Prevention, Brigham and Women’s Hospital
  • Professor of Medicine, Harvard Medical School
  1. Focus on Women
  2. Diagnostics requires women – large trials and power studies by gender
  3. CRT,
  4. Optimizing care for Women
  5. EF, CHF, MI are prevalent in Women
  6. Migraine in Women – related to CVD
  • EVP & Head, Global Commercial Development, Mylan
  1. Information on differences between women and men – Cholesterol
  2. Woman present with different symptoms – more progress because care is delayed
  3. Stable angina and zero plaque cardiac rehab
  4. Female specific guidelines
  5. wholistic approach, girl scouts as a start
  • CV Therapeutic Area Lead, Global Business Development, Pfizer
  1. Number of women in trials? 25% – how to extrapulate from this data?
  2. How to design trials, powering, endpoints, clinical trials, FDA – mandates reporting of Women representation in studies
  3. Data Gap – retrospective study – 30% women, guidelines based on 70% Men data
  4. Awareness – who is the PCP to close the Gap
  5. OBGYN is often the PCP, the only Annual a Women goes to
  6. Precision medicine in Women, what is actionable what is not
  7. Harness Phynotypic leverage repository
  • Medical Director, Boston Scientific
  1. Women vs Minority Women – Improvement will occur if tools and strategies will represent all demographic
  2. Accurate measurements, Women participation in trials, Latinos, Minority Women – not as % in the population
  3. best practices and guidelines
  4. Awareness, nosea and fatigue as symptoms,
  • Co-Director, MGH Heart Center Corrigan Women’s Heart Health Program, Massachusetts General Hospital
  • Associate Professor, Harvard Medical School
  1. Heart attacks in Women and prevention, awareness among women
  2. Impact of Pregnancy: Preeclemxia, HTN, DM, hematologic disease, small gestation newborn, Minority Pregnant women – diet
  3. 30% less referred for Aortic stenosis or transplantation
  4. Care for Patient vs Episodic Care
  5. Stress in Women – metric to measure in PCP
  6. AI to be used in referral based on Medical data
  7. Migrane – several medications need to be studied on Women with the disease
4:30 pm – 5:20 pm
Boston Scientific Ballroom
Disruptive Therapeutic Platforms: New Tools, New Outcomes

Recent advances of biological drugs have broadened the scope of therapeutic targets for a variety of human diseases. This holds true for dozens of RNA-based therapeutics currently under clinical investigation for diseases including heart failure. These emerging drugs could be considered in context of genomic/germ line screening, family history and epigenetics.

Moderator: Tony Coles, MD
  • CEO, Yumanity Therapeutics
  1. one of three death in the World
  2. Limb Ischemia
  • CEO, Moderna
  1. mRNA, clinical stage, published Human data Immuno oncology, VGEF therapeutics after MI
  2. Recombinant VGEF, PK goes to the heart mascle if goes to serum is degraded by nucleatase
  3. Post MI in pigs, Phase I, Phase II
  4. Chronic response formulation short half life (6 days)
  5. Step by step, get the right protein
  6. Cardiology – mRNA drug for one patient
  • CEO, Editas Medicine
  1. CRISPR technology – translational medicine changes in DNA
  2. viral vector therapy delivery: Eye liver, blood — easier for delivery
  3. Immune response from delivery of CRISPR molecule: control over the time response of the molecule: Immunogenicity
  4. Using biology knowledge
  • Center for Cancer Immunology, Massachusetts General Hospital
  • Member of the Faculty of Medicine, Harvard Medical School
  1. Cancer Immune response plays a role
  2. CVD and the Immune System: Transfer from Oncology to CVD: Mutations on genes mutations are not silent to the immune systems — development of Vaccine
  3. Oncologists  in lung cancer saw immune response against their own tumors
  4. macrophage in the heart
  • CEO, Alnylam
  1. CVD Program – Phase III
  2. PCSK9 – as a target genetically defined mutation, Hyper-cholesteronemia – subcutaneous delivery – Lowering LDL by bi-annual injection or quarterly – non-complaint with Statin
  3. ADVANCED medicine for CVD
  • Founder, AnGes
  1. Gene therapy – pipeline of 8  –
  2. DNA Vaccine for HTN
  3. Muscular therapy – Ischemia
  4. CVD – Reduction comorbidity and mortality
5:20 pm – 6:00 pm
Novartis Foyer

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Regulatory MicroRNAs in Aberrant Cholesterol Transport and Metabolism

Curator: Marzan Khan, B.Sc

Aberrant levels of lipids and cholesterol accumulation in the body lead to cardiometabolic disorders such as atherosclerosis, one of the leading causes of death in the Western World(1). The physical manifestation of this condition is the build-up of plaque along the arterial endothelium causing the arteries to constrict and resist a smooth blood flow(2). This obstructive deposition of plaque is merely the initiation of atherosclerosis and is enriched in LDL cholesterol (LDL-C) as well foam cells which are macrophages carrying an overload of toxic, oxidized LDL(2). As the condition progresses, the plaque further obstructs blood flow and creates blood clots, ultimately leading to myocardial infarction, stroke and other cardiovascular diseases(2). Therefore, LDL is referred to as “the bad cholesterol”(2).

Until now, statins are most widely prescribed as lipid-lowering drugs that inhibit the enzyme 3-hydroxy-3methylgutaryl-CoA reductase (HMGCR), the rate-limiting step in de-novo cholesterol biogenesis (1). But some people cannot continue with the medication due to it’s harmful side-effects(1). With the need to develop newer therapeutics to combat cardiovascular diseases, Harvard University researchers at Massachusetts General Hospital discovered 4 microRNAs that control cholesterol, triglyceride, and glucose homeostasis(3)

MicroRNAs are non-coding, regulatory elements approximately 22 nucleotides long, with the ability to control post-transcriptional expression of genes(3). The liver is the center for carbohydrate and lipid metabolism. Stringent regulation of endogenous LDL-receptor (LDL-R) pathway in the liver is crucial to maintain a minimal concentration of LDL particles in blood(3). A mechanism whereby peripheral tissues and macrophages can get rid of their excess LDL is mediated by ATP-binding cassette, subfamily A, member 1 (ABCA1)(3). ABCA1 consumes nascent HDL particles- dubbed as the “good cholesterol” which travel back to the liver for its contents of triglycerides and cholesterol to be excreted(3).

Genome-wide association studies (GWASs) meta-analysis carried out by the researchers disclosed 4 microRNAs –(miR-128-1, miR-148a, miR-130b, and miR-301b) to lie close to single-nucleotide polymorphisms (SNPs) associated with abnormal metabolism and transport of lipids and cholesterol(3) Experimental analyses carried out on relevant cell types such as the liver and macrophages have proven that these microRNAs bind to the 3’ UTRs of both LDL-R and ABCA1 transporters, and silence their activity. Overexpression of miR-128-1 and miR148a in mice models caused circulating HDL-C to drop. Corroborating the theory under investigation further, their inhibition led to an increased clearance of LDL from the blood and a greater accumulation in the liver(3).

That the antisense inhibition of miRNA-128-1 increased insulin signaling in mice, propels us to hypothesize that abnormal expression of miR-128-1 might cause insulin resistance in metabolic syndrome, and defective insulin signaling in hepatic steatosis and dyslipidemia(3)

Further examination of miR-148 established that Liver-X-Receptor (LXR) activation of the Sterol regulatory element-binding protein 1c (SREBP1c), the transcription factor responsible for controlling  fatty acid production and glucose metabolism, also mediates the expression of miR-148a(4,5) That the promoter region of miR-148 contained binding sites for SREBP1c was shown by chromatin immunoprecipitation combined with massively parallel sequencing (ChIP-seq)(4). More specifically, SREBP1c attaches to the E-box2, E-box3 and E-box4 elements on miR-148-1a promoter sites to control its expression(4).

Earlier, the same researchers- Andres Naars and his team had found another microRNA called miR-33 to block HDL generation, and this blockage to reverse upon antisense targeting of miR-33(6).

These experimental data substantiate the theory of miRNAs being important regulators of lipoprotein receptors and transporter proteins as well as underscore the importance of employing antisense technologies to reverse their gene-silencing effects on LDL-R and ABCA1(4). Such a therapeutic approach, that will consequently lower LDL-C and promote HDL-C seems to be a promising strategy to treat atherosclerosis and other cardiovascular diseases(4).


1.Goedeke L1,Wagschal A2,Fernández-Hernando C3, Näär AM4. miRNA regulation of LDL-cholesterol metabolism. Biochim Biophys Acta. 2016 Dec;1861(12 Pt B):. Biochim Biophys Acta. 2016 Dec;1861(12 Pt B):2047-2052


2.MedicalNewsToday. Joseph Nordgvist. Atherosclerosis:Causes, Symptoms and Treatments. 13.08.2015


3.Wagschal A1,2, Najafi-Shoushtari SH1,2, Wang L1,2, Goedeke L3, Sinha S4, deLemos AS5, Black JC1,6, Ramírez CM3, Li Y7, Tewhey R8,9, Hatoum I10, Shah N11, Lu Y11, Kristo F1, Psychogios N4, Vrbanac V12, Lu YC13, Hla T13, de Cabo R14, Tsang JS11, Schadt E15, Sabeti PC8,9, Kathiresan S4,6,8,16, Cohen DE7, Whetstine J1,6, Chung RT5,6, Fernández-Hernando C3, Kaplan LM6,10, Bernards A1,6,16, Gerszten RE4,6, Näär AM1,2. Genome-wide identification of microRNAs regulating cholesterol and triglyceride homeostasis. . Nat Med.2015 Nov;21(11):1290


4.Goedeke L1,2,3,4, Rotllan N1,2, Canfrán-Duque A1,2, Aranda JF1,2,3, Ramírez CM1,2, Araldi E1,2,3,4, Lin CS3,4, Anderson NN5,6, Wagschal A7,8, de Cabo R9, Horton JD5,6, Lasunción MA10,11, Näär AM7,8, Suárez Y1,2,3,4, Fernández-Hernando C1,2,3,4. MicroRNA-148a regulates LDL receptor and ABCA1 expression to control circulating lipoprotein levels. Nat Med. 2015 Nov;21(11):1280-9.


5.Eberlé D1, Hegarty B, Bossard P, Ferré P, Foufelle F. SREBP transcription factors: master regulators of lipid homeostasis. Biochimie. 2004 Nov;86(11):839-48.


6.Harvard Medical School. News. MicoRNAs and Metabolism.


7. MGH – Four microRNAs identified as playing key roles in cholesterol, lipid metabolism



Other related articles published in this Open Access Online Scientific Journal include the following:


  • Cardiovascular Diseases, Volume Three: Etiologies of Cardiovascular Diseases: Epigenetics, Genetics and Genomics,

on Amazon since 11/29/2015



HDL oxidation in type 2 diabetic patients

Larry H. Bernstein, MD, FCAP, Curator



HDL-C: Target of Therapy – Steven E. Nissen, MD, MACC, Cleveland Clinic vs Peter Libby, MD, BWH

Reporter: Aviva Lev-Ari, PhD, RN



High-Density Lipoprotein (HDL): An Independent Predictor of Endothelial Function & Atherosclerosis, A Modulator, An Agonist, A Biomarker for Cardiovascular Risk

Curator: Aviva Lev-Ari, PhD, RN



Risk of Major Cardiovascular Events by LDL-Cholesterol Level (mg/dL): Among those treated with high-dose statin therapy, more than 40% of patients failed to achieve an LDL-cholesterol target of less than 70 mg/dL.

Reporter: Aviva Lev-Ari, PhD., RN



LDL, HDL, TG, ApoA1 and ApoB: Genetic Loci Associated With Plasma Concentration of these Biomarkers – A Genome-Wide Analysis With Replication

Reporter: Aviva Lev-Ari, PhD, RN



Two Mutations, in the PCSK9 Gene: Eliminates a Protein involved in Controlling LDL Cholesterol

Reporter: Aviva Lev-Ari, PhD, RN


Artherogenesis: Predictor of CVD – the Smaller and Denser LDL Particles

Reporter: Aviva Lev-Ari, PhD, RN



A Concise Review of Cardiovascular Biomarkers of Hypertension

Curator: Larry H. Bernstein, MD, FCAP



Triglycerides: Is it a Risk Factor or a Risk Marker for Atherosclerosis and Cardiovascular Disease ? The Impact of Genetic Mutations on (ANGPTL4) Gene, encoder of (angiopoietin-like 4) Protein, inhibitor of Lipoprotein Lipase

Reporters, Curators and Authors: Aviva Lev-Ari, PhD, RN and Larry H. Bernstein, MD, FCAP



Excess Eating, Overweight, and Diabetic

Larry H Bernstein, MD, FCAP, Curator



Obesity Issues

Larry H. Bernstein, MD, FCAP, Curator



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Reversing Heart Disease: Combination of PCSK9 Inhibitors and Statins – Opinion by Steven Nissen, MD, Chairman of Cardiovascular Medicine at Cleveland Clinic

Reporter: Aviva Lev-Ari, PhD, RN

UPDATED on 2/25/2019

While nearly 10% of middle-age adults in China have high risk for cardiovascular disease, only 0.6% of these high-risk individuals use statins and 2.4% take aspirin, a national screening project reported in the Annals of Internal Medicine.

UPDATED on 5/5/2017

Europeans Mull PCSK9i Post-FOURIER Fallout on Clinical Practice

Patrice Wendling, May 04, 2017

But it was panelist Dr Stephen Nicholls (University of Adelaide, Australia) who took aim at the elephant in the packed auditorium. At an annual cost of about $14,100 for evolocumab and $14,600 for alirocumab (Praluent, Sanofi/Regeneron), the important question facing cardiologists is who will be eligible for these drugs “in a world where we can’t just write a scrip for every FOURIER-type patient; we won’t be allowed to.”

He suggested initially this will include patients with familial hypercholesterolemia and only those with established atherosclerotic CVD whose LDL-C remains unacceptably high despite therapy. Future FOURIER subanalyses may define other eligible high-risk groups.




UPDATED on 3/14/2017

PCSK9 Inhibitor Access Snarled in Red Tape, Rejections

Patrice Wendling, March 21, 2017

To determine whether this experience is happening nationwide, Navar and colleagues examined first PCSK9 prescriptions in 45,029 patients (median age 66 years; 51% female) between August 1, 2015 and July 31, 2016 in the Symphony Health Solutions database, which covers 90% of retail, 70% of specialty, and 60% of mail-order pharmacies in the US.

Nearly half (48%) of prescribers were cardiologists, and 37% were general practitioners. Most patients (52%) had government insurance, typically Medicare, and 40% had commercial insurance.

In the first 24 hours after being submitted to the pharmacy, 79.2% of prescriptions were rejected. Ultimately, 52.8% of all PCSK9 prescriptions were rejected.

Of special note, 34.7% of prescriptions for the pricy lipid-lowering drugs were abandoned at the pharmacy.



How 2 Drugs Lower Cholesterol Remarkably — and Reverse Heart Disease

Study shows promise for combination of newer drug and statins

How 2 Drugs Lower Cholesterol Remarkably --- and Reverse Heart Disease

Newer cholesterol-lowering drugs combined with more conventional medicine reduces bad cholesterol to incredibly low levels, a new study shows. Perhaps even more important, the combination also reduces the heart-attack-inducing plaque that forms inside the arteries, the study says.

The study was led by cardiologist Steven Nissen, MD, Chairman of Cardiovascular Medicine at Cleveland Clinic. Results appeared recently in the Journal of the American Medical Association (JAMA).

The study looked at the use of a drug called evolocumab by people who took statins to lower the amount of LDL, or bad, cholesterol in their blood. Evolocumab is a drug called a PCSK9 inhibitor. This is a newer kind of medicine that can make LDL cholesterol levels plummet.

The people who took statins and evolocumab had greater reductions in atherosclerosis compared with people who took statins and a placebo.  Atherosclerosis is  a disease in which plaque builds up inside your arteries.  The condition can lead to serious problems, including heart attack, stroke, or even death.

The results are an intriguing indicator — rather than definite proof — that evolocumab may have benefit for patients taking statins, Dr. Nissen says. Researchers are still awaiting the results of large clinical trials investigating whether evolocumab is safe and will prevent heart attack, stroke or death. The first results of these studies are expected in April 2017.

Special ultrasound

In the study, researchers treated for 18 months 968 high-risk people who had extremely high levels of blood cholesterol.

Participants were randomly assigned to take either a statin and a placebo, or a statin and evolocumab.

Researchers monitored the participants’ cholesterol levels. They also used a special ultrasound probe to measure the amount of plaque in their arteries at the beginning and the end of the study. 

“We were able to show that getting the bad cholesterol levels down to really low levels, down to the 20s and 30s, can actually remove plaque from the coronary arteries,” Dr. Nissen says. “This going to levels that we’ve never been able to achieve before.”           

Low cholesterol, less plaque

Results show the group treated with statins and a placebo reduced their LDL cholesterol levels to 93 on average. At the same time, the group treated with the combination of the statin plus evolocumab got down to an average bad cholesterol level of 36.6.

“No one’s ever reached levels that low in a clinical trial,” Dr. Nissen says.

Participants who took evolocumab also had less plaque in their arteries at the end of the study — essentially reversing their heart disease.

“We, for the first time now, have shown that this new class of drugs, the PCSK9 inhibitors, has a favorable effect on the development of plaques on the coronary artery and can actually regress those plaques,” Dr. Nissen says. “And it turns out about two-thirds of patients actually had less plaque at the end of 18 months than they started with.” 

PCSK9 inhibitors, which are expensive, are not for everybody, Dr. Nissen says. Currently, the drug is used in addition to statins for the highest-risk patients with particularly high cholesterol.


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LIVE 9/21 8AM to 2:40PM Targeting Cardio-Metabolic Diseases: A focus on Liver Fibrosis and NASH Targets at CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston






Nonalcoholic Steatohepatitis (NASH)


Leaders in Pharmaceutical Business Intelligence (LPBI) Group is a

Media Partner of CHI for CHI’s 14th Annual Discovery on Target taking place September 19 – 22, 2016 in Boston.

In Attendance, streaming LIVE using Social Media

Aviva Lev-Ari, PhD, RN




Wednesday, September 21

7:30 am Registration Open and Morning Coffee

8:00 Chairperson’s Opening Remarks

Rebecca Taub, M.D., Ph.D., CEO, Madrigal Pharmaceuticals

  • Epidemic of NASH,
  • approaches to treating NASH – Fibrosis
  • NASH is a metabolic Disease of the Liver
  • Treating the HCV will treat the Fibrosis

8:10 FEATURED PRESENTATION: The Epidemic of Fatty Liver Disease: Silent, Serious and Still Growing?

Lee Kaplan, M.D., Ph.D., Director, Obesity, Metabolism and Nutrition Institute, Massachusetts General Hospital, Harvard Medical School

  • Silent, Serious and Growing
  • Obesity the Disease = BMI>30: Medical Complicastions for BMI >%) – On ANti-Obisity and Bariatric SUrgery, Type 2 Diabetis .. NAFLD .. NASH .. Cirrhosis .. HCC
  • Parkinson’s Disease
  1. Medical Complications of Obisity =197 :
  2. NAFLD – Nonalcoholic Fatty Liver Disease >>> Liver transplantation replacing HCV
  3. Associated with obesity and type 2 diabetes
  4. NAFLD is UP 90% wiht Severe Obesity
  5. Viral hepatitis and Hemochromatosis
  6. NAFLD: Steatosis, Inflamamtion, Hepatocellular Necrosis, Fibrosis, Cirrhosis
  7. NASH: insulin resistence .. metabolic syndrom .. interaction
  8. Alternative Model: Metabolis Syndrom.. Steatosis .. NASH … FIbrosis
  9. Genetics of Liver DIsease
  10. PNPLA3 Associated with NAFLD – Not Weight Gain
  11. Other genes: A Partial List:
  12. Diagnosis of NASH: Liver biopsy macrovescicular fatty change: InflammationMollery bodies
  13. 75% Patients with Cirrhousis have obisity
  14. Alcoholoc hepatisis >> Progression to Cirrhousis
  15. Macrovesicular Steatosis
  16. NASH – inflammation
  17. Sinusoidal Pericellular Fibrosis –
  18. LAB Features of NAFLD
  • Transaminase elevation
  • Akaline phosphate
  1. Biomarkers – NASH – associated cirrhousis with lower rate 30% of elevation
  2. Fibrosure
  • Clinical Features of NASH: none presentation, Bright, Echo Fibroscan FibroscanScreen for HCC, Varices if Gray zone: Biopsy
  • Treatment of NASH
  • Treat liver disease: Treat steatosis then Inflamamtion and fibrosis
  1. NAFLD Treatment Strategy: Stepwise Approach
  • Treat the steatosis Piodlitazone
  • PPARalpha, delta,
  • Treat Inflammation: ANtioxidant
  • CCR2/CCR% inhibitors
  • Metabolic SUrgery
  • Weigh-independent for bariatric
  • Bariatic: improvrment of steatosis,effect on inflammationless clear
  • dramatic on weigh loss
  • NO clear is surgery improved cirhousis
  • If NASH developed >>>> progression s the rule
  • No great treatment of NASH

Medication-assciated NASH: Glucocorti


8:40 Non-Alcoholic Steatohepatitis and Cardiovascular Disease: Modulation by Novel PPAR Agonists

Bart Staels, Ph.D., Professor, INSERM, University of Lille, Pasteur Institute

Peroxisome proliferator-activated receptors (PPARs) are ligand-activated nuclear receptors which regulate lipid and glucose metabolism as well as inflammation. In this presentation, we will review recent findings on the pathophysiological role of PPARs in the different stages of non-alcoholic fatty liver disease (NAFLD), from steatosis development to steatohepatitis and fibrosis, as well as the preclinical and clinical evidences for potential therapeutical use of PPAR agonists in the treatment of NAFLD. PPARs play a role in modulating hepatic triglyceride accumulation, a hallmark of the development of NAFLD. Moreover, PPARs may also influence the evolution of reversible steatosis towards irreversible, more advanced lesions. Large controlled trials of long duration to assess the long-term clinical benefits of PPAR agonists in humans are ongoing.

Non-Alcoholic Steatohepatisis and CVD – Meta inflammatory disease

  • NAFL — abnormal Lipid accumulation
  • NASH >> Balooning, FIbrosis inflamation
  • Resolution of NASH is associated with reduction of Fibrosis (Golden – 505 trial)

CVD is linked to NAFLD: Lipids elevated and therosclerosis

  • TG – elevated, APO B elevated, VLDL – elevated HDL decrease
  • PPAR Alpha
  • Gamma
  • PPAR Beta/Delta agonist: GENFIT – Elafibranor

Trans-activation: Lipid and Glucose homeostasis: Trans-repression – anti-inflammatory properties

  • Hapatic mitochondrial activity deseases upon progression from NAFL to NASH: Obese NAFL and NASH
  1. Upregulated hepatic respiratory in obese humans with or without NAFL
  2. Impaired
  3. Hepatic PPARalpha Expression Decreases upon Progression of Nash and Fibrosis
  4. hepatic PPARalpha expression – target genes increase in patients with improved NASH histology after 1 year
  5. Metabolic Regualtion by thehepatic JNK Signaling Pathway
  6. Target gene transcription – miR-21 expression increases in human
  7. PPAR Delta: Elafibbranor: – effect on plasma lipids: A Dual PPAR alpha/Delts (GFT505): 80mg vs placebo and 120mg vs placebo, improves plasma apolipolipids and glucose HbA1C – insulin sensitivity
  8. efficacy in NASH acting on: Steatosis, fibrosis and cirrhosis
  9. inflammatory markers: RESOLVE-IT Phase 3 Study Desing: NASH ressolution without adverse on FIbrosis and Cirrhosis

GOLDEN505 Trial: Improves plasma lipid levels: Triglycerides

Inclusion Criteria:


Improve atherogenic dyslipidemia

  • APOC3 – associated with CVD

9:10 PANEL DISCUSSION: Liver Fibrosis and NASH Targets

Moderator: H. James Harwood, Ph.D., Delphi BioMedical Consultants, LLC


Lee Kaplan, M.D., Ph.D., Director, Obesity, Metabolism and Nutrition Institute, Massachusetts General Hospital, Harvard Medical School

Bart Staels, Ph.D., Professor, INSERM, University of Lille, Pasteur Institute

Rebecca Taub, M.D., Ph.D., CEO, Madrigal Pharmaceuticals

Weilin Xie, Ph.D., Senior Principal Scientist, Biotherapeutics, Celgene Corp.

  • FDA’s view on surrogate endpoints
  • Biomarkers of NASH
  • Regulatory challenges
  1. Liver biopsy: gold standard, invasive direct measure of endpoints pros/cons
  2. non-invasive functional tests – plasma bioamrkers
  3. non-invasisve liver imaging techniques: MRI to assess hepatic fat content MRE to assess hepatic fibrosis, Fibroscan,
  4. Endpoints acceptable by FDA: Current vs Future
  • Pre clinical Translational animal models

Discussion by Panel members

Progression from NAFLD to NASH: Oxidative stress and toxic lipids

NASH and Steatosis are different populations

Alcoholoc Steatosis vs Non-Alcoholic Steatosis

  • Obesity cause of Fatty liver
  • NASH in Diabetes
  • NASH progresses
  • Steatosis is associated with NASH
  • Different types of NASH: HTN, Dislipedemia,
  • GENETICS underlining factors, more genes are discovered
  • Limitations of Animal Studies for inference on Humans – careful in over generalizing results
  • Metabolic SYndrom -not all progresses to NASH
  • Nonalcoholic Steatohepatitis (NASH) depend on Steatosis


9:40 Coffee Break in the Exhibit Hall with Poster Viewing

10:25 Targeting Fibroblast Activation Protein (FAP) and FGF21 to Treat Fatty Liver Disease

Diana Ronai Dunshee, Ph.D., Department of Molecular Biology, Senior Scientific Researcher, Genentech, Inc.

FGF21 is a hormone with anti-obesity and hepatoprotective properties. However, the beneficial effects of FGF21 are limited by a relatively short half-life in circulation. We discovered that fibroblast activation protein (FAP), an endopeptidase overexpressed in liver with cirrhosis, cleaves and inactivates FGF21. Pharmacological inhibition of FAP increases endogenous levels of active FGF21, thus making FAP a promising target for the treatment of non-alcoholic-steatohepatitis (NASH).

  • Medical complications of obisity: NASH and DM-2
  • energy consumption
  • white adipose tissue – energy storage
  • brown adipose tissue matochondia’s energy
  • FGF21 – Human activation of protein cleavage: A Homone beneficial on metabolic health circulation, weigh loss
  • it suppreses hepatic Steatohepatitis
  • One singleinjection in mice — leads to energy expenditure induced weigh loss and metabolic improvement in Obese Humans
  • Negative FGF21 is Rapidly Eliminated from the body – renal degradation and Inactivation of FGF21 Endopeptidase Cleavage Site – Fibroblast Activation Protein Matched FAP Endopeptidease Specificity
  • Closest relative of DPP4 upregulted during tissue injury in NASH
  • FAP is SUfficient to Cleave FGF21
  • Recombinant FGF21 with Recombinant FAP in Serum or Plasma
  • FAP Protease – Serum Immunodepleted Ablates FGF21 Cleavage Activity: Peptide IgG vs anti-FAP
  • FAP Cleavage Inactivates Human FGF21 dependent on KLB-FGFR1c placed on the site
  • hFGF21 in Not Cleaved in FAP KO Mice
  • Fc-hFGF21 is more stable in FAP KO mice
  • FAR cleaves Endogenously Produced FGF21 In Vivo in monkeys and in dogs
  • The FAP Cleavage Consensus GLY-Pro is COnserved in most mammalian FGF21
  • FAP Does not Cleave the C-Terminal Residues of Mouse FGF21
  • Human: FAP, DPPIV
  • Mouse: FAP, DPP4
  • FAP INhibition
  • FAP is Overexpressed in Liver with Steatohepatitis: Early NASH vs Late NASH
  • Proposal: FAP Inhibition for FGF21 Stabilization in NASH
  1. Fatty hepatocytes – e.g. NASH
  2. Activated stellate cells, e.g. NASH


10:55 Thyroid Hormone Receptor Beta (THR-ß) Agonist for NASH: Correcting a Primary Deficiency in NASH Livers

Rebecca Taub, M.D., Ph.D., CEO, Madrigal Pharmaceuticals

NASH patients typically have metabolic syndrome including diabetes, dyslipidemia, obesity, and primarily die of cardiovascular disease. Hypothyroidism at the level of the thyroid gland and liver-specific hypothyroidism are common in NASH. Based on clinical and preclinical data, Thyroid receptor beta agonists decrease insulin resistance, reduce LDL-C, triglycerides fatty liver, inflammation and fibrosis in NASH. The target will also provide CV benefit to patients with NASH. MGL-3196 is a highly THR-ß selective liver-directed once daily oral medication that has shown excellent safety and lipid-lowering efficacy in humans; unlike prior thyroid receptor agonist(s), no cartilage findings in chronic toxicology or ALT increases in human studies. MGL-3196 is being advanced in Phase II studies in patients with genetic dyslipidemia or NASH.

Madrigal Portfolio of drugs:

  • MGL-3196: First-in-Class THR-Beta Agonist – discovered first at ROCHE – THR-beta selective targeted to the Liver – regulated by THR-Alpha  – in Phase II – no side effects on bone
  • Large & underserved Markets in NASH
  • Phase 2 HeFH Patients
  1. Hypothyroidism common in NASH patients
  2. Liver-specific Hypothyroidism present in human NASH degradation of thyroid hormone increases deiodised 9DIO) 3 produced by Stelllate cells in NASH liver
  3. Treating NASHrather than fibrosis is key in addressing the disease – approvable endpoint
  4. THR – Thyroid hormone reduces Cholesterol
  5. Thyroid hormone T3 thyroxine – treatment amy cause osteoporosis
  6. MGL 0 3196: Liver size, Live Triglycerides, Improve Insulin tolerance, decrease ALT
  7. Reduction of key NASH, Fibrosis Pathway Genes at Human Comparable Drug levels
  8. THR-beta: Decreased Liver Fibrosis, Apoptosis in mice:


  • Single ascending dose study
  • Multiple – ascending studies: LDL and TG decrease
  • decrease Non-HDL CHolesterol
  • Decrease Apolipoprotein B
  • Pleiotropic Pioglitazone Effect in NASH at 6 month treatment and biopsy of liver – dramatic effect in NASH – ten years ago study
  • PPAR gamma agonist – NEGATIVE SIDE EFFECTS: weight gain, CHF, Bone osteoporosis
  • Anti-inflammatory: well tolerated

No Single NASH Therapeutics – Conbination agents

MGL – 3196 Phase 2 – Study: Proposed Phase 2 Proof of COncepts NASH Protocol

  • Unmet needs in FH, a severeGenetic Dyslipedemia
  • Weight loss in 6 weeksreduction in cholesterol and TG
  • Likelihood of Success
  • second study after 9 months
  • is different on NASH Patients in 12 weeks using MRI on Liver
  • prevalence
  • HeFH, PCSK9 inhibitors plus standard care
  • Unique and Complementary Lipid Lowering Profile
  1. Lowers Lp(a) and severely atherogenic practice
  2. Proposed Phase 2 HeFH Patients


11:25 Enjoy Lunch on Your Own


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LIVE 9/20 8AM to noon GENE THERAPIES BREAKTHROUGHS at CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston






Leaders in Pharmaceutical Business Intelligence (LPBI) Group is a

Media Partner of CHI for CHI’s 14th Annual Discovery on Target taking place September 19 – 22, 2016 in Boston.

In Attendance, streaming LIVE using Social Media

Aviva Lev-Ari, PhD, RN



COMMENTS BY Stephen J Williams, PhD

Gene Therapy Breakthroughs

New Strategies for Better Specificity and Delivery


2:05  Chairman’s Remarks

Joseph Gold, Ph.D. Director Manufacturing Center for Biomedicine and Genetics, Beckman Research Institute City of Hope


  • CBG (center for biomedicine and Genetics) 20000 sq feet
  • CTPC (center therapy production) mainly CART
  • CBG 16 years operation do all stem cells and >400 products
  • New stem cell Beta cell progenitor
  • Do oncolytic VSV
  • CTPC is investigator driven CART islet cells,
  • Like to do novel work so work with CIRM
  • Banking of modified stem cells
  • Adherent scale out limitations: cost,inefficient; solution can be suspension
  • Establish hESC; plate on CELLstart > Accutase>StemPRO SFM>differentiation process; defined reagents — they use this for cardiomyocyte differentiation: they are functional (inotropy, chronotropy response to isoproterenol) can freeze back cells
  • Create a bank of intermediate cells and when you need it for surgery they will put on their matrix, enrich, expand and ship out
  • Allogeneic cells: project where take allogeneic neural stem cells to deliver a chemotherapy payload as they like to migrate to brain tumors
  • Allogeneic cells: for ALS modified to express GDNF
  • HIV resistance with engineered CCR5 negative blood stem cells
  • Release assay considerations: viability, sterility, if cryopreserved then can determine identity, viral insertions, VSV-G copy number, endotoxin and potency (FDA is wanting phase I potency assays) for CART potency is % transduced
  • Good in vivo activity of the neural stem cells loaded with chemotherapeutic



  • If deliver GDNF to muscle  using genetically modified myoblasts
  • Best to use fetal stem cells – less issues


Canavan disease: progressive fatal neurologic disorder that begins in infancy and don’t make it past teenage years

  • Rossbach is taking autologous cells reprogramming generating iPS cells and then modifying by CRISPR but the CRISPR issues of off target effects persist as well the time required for process and verification; also don’t want to use a selectable marker and put in patients; so you can differentiate the cells and hit them with a lentiviral vector system


They have been named a PACT Center Production Assistance for Cell Therapy where you can apply for a project grant.  Applicable for startups up to larger mature companies



They do a standard panel of tests for viral infections.

They work with investigators or companies at all stages of manufacturing processes.





2:15 Large-Scale Production of Cell Therapies for Regenerative Medicine

Joseph Gold, Ph.D. Director Manufacturing Center for Biomedicine and Genetics, Beckman Research Institute


2:45  Directed Evolution of New Viruses for Therapeutic Gene Delivery

David Schaffer, Ph.D.  Professor of Chemical and Biomolecular Engineering, BioEngineering, Molecular and Cell Biology and Neuroscience;


AAV is very safe as many people already infected with it


  • Spark (Leber’s cogenital anaurosis
  • Hemophilia B
  • Lipoprotein lipase deficiency
  • Spinal muscular atrophy
  • Challenges: are we just getting the ‘low hanging fruit’ eg Spark therapy must be injected after retinal therapy, hemo B needs to be given at high doses
  • Their theory was AAV had been evolving for its own purposes so hence the limitations of AAV;
  • Utilized 25 different techniques to generate variants of AAV in a library then packaged (each will have its own barcode)
  • Broad platform technology: retina, lung, brain and spinal cord

Retinal:  AAV may be too large to get through layers of the eye, problems; subretinal injections and damage or retinal detachment.  Then they used their whole library in an in-vivo screen (as hard to recapitulate the multi cell layers of the retina).  


Cystic Fibrosis

  • AAV2H22 variant worked very well to supply the CFTR gene in pig model of cystic fibrosis and increases chloride transport and reduce bacterial load
  • Then found pig variant AAV did not work well on human tissue so designed a human variant and worked well in human tissues
  • The variant AAV2.5T surrounds sialic acid binding pockets and increases binding and endocytosis


Brain and Spinal Cord:  Sanfilippo B trial  8 holes drilled into skull followed by 16 AAV injections


  • Injected a generated AAV variant (by evolution process) : engineered AAV2 is 100 fold better getting through blood brain barrier… novel variant undergoes retrograde transport to cortex ; made a cas9 to remove a tdTomato gene overexpressed in mouse and found 90% knockdown
  • Also interesting point: the porcine variant did not work in human and the human variant did not work in porcine.  Implication for FDA safety and efficacy testing must do in monkeys

They started a spinout 4D Molecular Therapeutics


4:25 Lentiviral Vectors for Gene Therapy


Munapaty Swani, Ph.D. Texas Tech Health Science Center


  • Can express multiple shRNA under a separate promoter but toxic so if expressed in miRNA backbone could be safer under a pol II
  • How much of flanking sequence is needed?
  • 30 nt flanking sequence is enough for Drosha processing
  • Constructed 1 to 7 shRNA-miR targeting CCR5 and 6 viral genes; all constructs were functional
  • Problem with pol ii promoter
  • These 7 shRNA miRNA protect against HIV entry if against CCR5 and the 7 viral elements
  • Used the non-integrating lentivirus for transient to see if infect T cells or not versus integrating lentivirus ; results non-integrating lentivirus did not infect t cells so safer to use
  • CCR5 disruption reduced HIV infection in T cells in vitro;
  • ZFN treatment of HIV+ PBMC prevents activation of HIV
  • Encapsulted CAS9 within LV; cas9 protein is incorporated within LV and is functional
  • First transduce then come in with the Cas9 so made all in one lentivirus with Cas9 and an sgRNA expression vector *******
  • This shows that it is possible to put all in a nanoparticle based lentivirus and an all in one may make it easier and safer (supposedly)


4:55 AAV Capsid Design

Miguel Seria Esteves, PhD Associate Professor Neurology, Gene Therapy Center University of Massachusetts Medical School


-AAV replication dependent no known human disease with native AAV

  •  Multiple barriers to get across blood brain barrier
  • AAV9 preferentially target neonatal neurons and adult astrocytes
  • Multiple capsids can be used for AAV9 infection in brain but not complete
  • Can we design better capsids to give it better tropic properties and better penetration to blood brain barrier
  • Using a polyalanine in the 5’ end of the caspid was most efficeint
  • Increases gene transfer efficiency especially IN SELECT CELL TYPES; Glial transduction and increased in striatum: increase is structure specific so little in thalamus but good in cerebellum and spinal cord
  • AAV9 tranduces also in peripheral tissues with or without modified capsid


Huntington’s Disease

  • Polyglutamate disease polyy glu on huntingtin protein
  • They get a 40 to 50% reduction of huntingtin but not significant between capsid design
  • They did a directed evolution of AAV capsid and generated capsid gene delivery diversity: DNA shuffling and in vivo selection
  • AAV-B1 is a new tropic capsid showing transduction of different structures
  • Five fold reduction in tropism to the liver but massive increases in muscle and beta exocrine cells and lung
  • Presence of neutralizing antibodies is a problem with AAV therapy
  • In conclusion unknown mechanisms by whivh a highly hydrophobic string of 19 alanines modifies the CHS tropism of AAV9 kvariants
  • Chimeric capsids identified from in vivo screen can reveal interesting patterns of tropism

8:20 AAV for Gene Therapy and Genome Editing

James Wilson, M.D., Ph.D., Professor, Department of Pathology and Laboratory Medicine, Perelman School of Medicine; Director, Orphan Disease Center and Director, Gene Therapy Program, University of Pennsylvania

AAV delivery for CNS can be direct into brain or into CSF but AAV are big and vectors usually don’t cross BBB

Mucopolysaccharidoses a lysosomal storage disease including Hunter, Morquio

  • There are canine models for these diseases
  • Data show that intrathecal delivery has good distribution to the CSF not the serum
  • With IV you only get spinal distribution but IT you get good distribution to cerebellum and frontal cortex
  • Intrathecal superior for clearing lesions in cortex of dogs
  • In the dog the disease is more skeletal and less neurologic so with IT dogs were better than control but still some problems
  • Avaxis: AAV9 gene therapy for SMA and ALS but trials are very small but seems to be dose dependent effect ; phase I/II done; Pfizer, Esteves and REGENEXBIO have trials
  • Liver transduction of AAV has always been a success; early vectors were not efficient for uptake but AAV9 hemophelia B (factor 9) was
  • OCTD disease of urea acid cycle (neonatal versus late onset); proteins altered metabolism
  • Have to create a metabolic sink to detoxify metabolites; not lie a gene replacement therapy
  • They got stable expression in mouse adult  model of OCTD with AAV and had rapid onset of expression; but when done in newborn mice they saw transient expression
  • Newborn liver is proliferating so gene vector may be diluted out versus the adult liver
  • So turned to gene editing (ZFN:NEHJ, TALEN:HR gene correction, CRSPR:transgene addition by HR
  • Staph aureus cas9 is smaller than most and can fit in a 4.79kb vector*****
  • Put in 2.6 kb doner OCTD gRNA
  • With a CRSPR-Cas9 mediated deliverycould maintain the expression of OCTD for over a week in newborn mice
  • BUT in adults the gene corrected animals started to die; they were losing their ability to break down protein
  • In newborns you got 10% gene editing with 30% indels but with adults 30% resulted only in 1% gene editing
  • There is a propensity to create large (>50bp) indels in the adult
  • NGS was needed to fully detect the target transgene integration, PCR is not good enough
  • Says that large animal models are needed for safety/efficacy studies
  • Problem with Rhesus monkey: started with a humanized mouse in Rag mice because did not want to do monkey studies; but did not get good expression in the monkeys (it was not the vector which was the problem)
  • AAV may be good enough of a donor to cause the HR recombination


Summary:  AAV vectors combined with a CRSPR CAS9 system is effective in neonatal delivery however 1) AAV by itself may be a good delivery system by itself but need the crsipir guide RNA to make the break to promote HR and get the best efficiency of integration 2) use of CRSPR CAS9 may direct the proper integration you want to deliver the OCTD exons to correct gene loci


Talk specific @ and #




9:20 Using CRISPR/Cas9 to Target and Destroy Viral DNA Genomes; Inactivating HBV


Bryan R. Cullen, Ph.D., James B. Duke Professor of Molecular Genetics and Microbiology and Director, Center for Virology, Duke University


The CRISPR array is an evolutionary record of the bacteriophage that the bacteria have encountered.

  • Incredible that a bacteria that never encountered a chromosomal structure would scan the genome with these gRNAs and then initiate HR and NEHJ (error prone in mammalian cells usually 3 nt
  • HBV is a very confused retrovirus because it first goes into the nucleus then becomes a template for RNA synthesis to make the particles that reinfect the cell and invisible to immune response
  • Although they do not produce infectious virus it still remains in genome
  • sgRNA screening: use a luciferase based assay to correct or knock out luc so looking for decrease of luciferase activity
  • In a model of HBV infection where they have an inducible HBV cccDNA in a single integrated copy the cas9 reduced the DNA but protein was not decreased that much (if you hit episomal DNA you see loss and the disintegration of cut out DNA but if you hit integrated DNA you see repair
  • In their case the integrated DNA was just mutated (A or AA insertion) so in essence a frameshift mutation
  • Future strategies: use two guide RNAs to permit deletions or allow use of nickase. Currently these gRNA use polIII which is very large
  • They are editing the VEGF loci using Sau Cas9 and two sgRNAs
  • In mice with HBV integrated in their genome get some cutting but they need to go to higher doses of Cas9 system
  • Potential future success if reduce viral load as HBV continually release antigen which results in T cell anergy and if reduce the viral load may help to reduce anergy and wake up the immune system


Meeting specific # and @






10:35 Targeted Endonucleases as Antiviral Agents: Promises and Pitfalls

Keith R. Jerome, M.D., Ph.D., Member, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center; Professor and Head, Virology Division, Department of Laboratory Medicine, University of Washington


  • Can cure Hepatititis C because can stop replication
  • HSV in neurons and live for life: long lived form like HBV
  • Herpes simplex (HSV) establishes in dorsal root ganglion: acyclovir might be useful but came off patent
  • He reached out to advocacy groups to get data on need for HSV cure to convince funding agencies this is important
  • They found it is important to people; 90% want a cure if 5 years away
  • Homing endonucleases: small 800bp high specificity easy to put in vectors more difficult retargeting to other DNA targets
  • www.ltk.uzh.ch/de/dyn
  • HSV homing endonuclease from Cellectis AG targets a 24bp sequence in Ul19; introduces a DSB at target site with 4 bp 3’ overhang;
  • AAV targeted endonuclease delivery; nonimmunogenic; persists in episomal state
  • Exposure to HSV specific HE decreases virus production from neuronal cultures at all stages of replication cycle; if infect and let go for a month but the HE disrupts HSV in acute late-acute and late cycle
  • Developed a mouse model of HSV infection and AAV delivery in vivo system; so the AAV was accessing the nerve endings and going down to the trigenital dorsal ganglia; transport is independent of AAV serotype but transgene expression is HIGHLY dependent on AAV serotype
  • The in vivo HE treatment appears well tolerated however just a casual observation
  • Used NGS with bioinformatic approach to determine off target sites for the most likely mouse loci
  • HE suppresses viral reactivation (used PCR based reactivation assay)


Talk Specific @ and #








12:05 pm CRISPR/Cas9 for the Screening of the Human Kinome – A Pilot Study in an Aggressive Pediatric Cancer Cell Line

Simone T. Sredni, M.D., Ph.D., Research Assistant Professor, Neurological Surgery, Northwestern University Feinberg School of Medicine, Ann and Robert H. Lurie Children’s Hospital of Chicago


LentiArray CRISPR Kinase Array

  • Malignant rhabdoid tumors (MRT); among most aggressive of pediatric tumors rare but lethal
  • Inactivating mutations in SMARCB1 (INI1 gene)
  • Component of swi/snf chromatin remodeling complex
  • Can originate anywhere in kidney brain and spine
  • Kinase inhibitors may be effective (PLK1, ERBB2, AURAKA) : AURKA inhibitor in phase 2 and giving promising response
  • Used LentiArray viral vector system and stable expressed Cas9
  • Tested 160 kinases in screen; high transfection efficiency
  • PIMs; protooncogenes (proviral common integrations sites in Maloney leukemia; overexpressed in prostate  and hematologic; effects cell cycle cdc25a is a target and cell survival targets as well; PIM3 high copy number in MRT as well as PIM2
  • KO limits proliferation increases senescence and confirmed by MTT with pan PIM inhibitor CS6258 (Cyclene Pharmaceuticals)
  • PLK4 overexpression in peds haploinsufficient mice do make tumors; is it mutated? Yes inactivating mutations
  • PLK4 expression higher through cell cycle – dependent?
  • KI67 was down with Knockdown
  • Decrease in clonogenic assay on plastic not soft agar
  • CFI-400945 is PLK4 inhibitor is ovarian trials
  • Is effective in MRT and comparable to their cas9 knockdown










Tuesday, September 20

7:00 am Registration Open and Morning Coffee



8:05 Chairperson’s Opening Remarks

Bryan R. Cullen, Ph.D., James B. Duke Professor of Molecular Genetics and Microbiology and Director, Center for Virology, Duke University

8:20 AAV for Gene Therapy and Genome Editing

James Wilson, M.D., Ph.D., Professor, Department of Pathology and Laboratory Medicine, Perelman School of Medicine; Director, Orphan Disease Center and Director, Gene Therapy Program, University of Pennsylvania

In vivo delivery of nucleic acid therapeutics remains the primary barrier to success. My lab has focused on the use of vectors based on adeno-associated virus (AAV) for achieving success in pre-clinical and clinical applications of gene replacement therapy. Most of the current academic and commercial applications of in vivo gene replacement therapy are based on endogenous AAVs we discovered as latent viral genomes in primates. These vectors are reasonably safe and efficient for application of gene replacement therapy. The emergence of genome editing methods has suggested more precise and effective methods to treat inherited diseases in which genes are silenced or mutations are corrected. AAV vectors have been the most efficient platform for achieving genome editing in vivo. We will review our attempts to achieve therapeutic genome editing in animal models of liver disease using AAV.

  • AAV for delivery of vector to CNS
  • Novel AAV Platform – capsid platform – distributed by PENN Vector
  • direct
  • IV
  • into CNS: Head and Spinal MPS – Mucopolysaccharidosis – Class of lyposomal Storage Disorder
  1. AAV9: Binds Glycans with Terminal Gal – BBB

CNS gene transfer in canine model of MPS VII following IV and intrathecal AA( administration

  • Serum
  • CSF: Frontal cortex, Cerebelum, Spinal Cord
  • Intravenous (IV) and Intracisternal (IC) – AA9

Gene Therapy for Motor Neuron Disease: SMA and ALS: PhaseI/II clinical trial of AAV9-SMN IV in infants with SMA1 – Nationwide Children’s

  • High dose +23.3 Month survival


  • Avexis
  • Abeona
  • Pfizer
  • Nationwide
  • Esteves

Liver Transduction Following IV Adm of AAV8 Vectors

  • Mouth liver Day 3 vs Day 90
  • Hemophilia B: Therapeutic protein

Urea Cycle Disorder

  • newborn OTCD infant in HA crisis

Goals: efficient but Transient vorrection following AAV* Gene Therapy in Newborn

transfer of caspid – promoter vector  to correct diffect

  • Survival: neonatal gene therapy – two doses of vector, three injections – immunogenic
  • Liver transplantation of neonatal before 1 year of age

Gene Editing

  • gene targeting by ZFNs, TALENs or CRISPR/Cas9
  • WT donor DNA
  • Transgene donor DNA
  • Gene disruptions

In Vivo correction – liver mouse by AAV. CRISPR-SaCas9

  • OTC donor template
  • efficient restoration of OTC Expression in the liver  – mice treated at Neonatal Stage by AAV8.CRISPR-SaCas9 – Vector administration
  • Cas9 – Kinetics of Cas9 – Week 1,3,8 – dilutes with time
  • Should work in Adult mice vs Neonatal mice: Low dose vs High dose
  • Ureogensis increased
  • On-target Deep Sequensing and their Distribution in Neonatal-treated and Adult-treated Animals

In vivo gene editing: mixed results: Site-directed Insertion of hOTCco Gene Cassette in the OTC gene : Controls: WT and spf(ash)

Western Blot: NGS analysis demonstrates on target transgene integration 20 to 32%

High Protein DIet to Evaluate the Efficacy

  • NEW BORN – Gene Targeting in FIX-KO Mouse by CRISPR/Cas9: Infron1, Exon 2 Infron2 Exon 3
  • ADULTS – Gene Targeting in FIX-KO Mouse by CRISPR/Cas9:

Gene therapy must accumulate experience in Animal models: Safety and Efficacy

NEGATIVE RESULTS IN MONKEYS: Analysis of Liver Tissue for Editing and SaCas9

  • CRISPR/Cas9 -mediated Gene knock down of rhPCSK9 in Monkeys (Rhesus Macaque) in LIVER
  • In vitro sgRNAs in monkeys and human cells
  • EGFP sgRNA vs hrPCSK
  • In vivo does not infer In vitro
  • Gene Editing in Human — we are not yet there

9:20 Using CRISPR/Cas to Target and Destroy Viral DNA Genomes

Bryan R. Cullen, Ph.D., James B. Duke Professor of Molecular Genetics and Microbiology and Director, Center for Virology, Duke University

A number of pathogenic human DNA viruses, including HBV, HIV-1 and HSV1, cause chronic diseases in humans that remain refractory to cure, though these diseases can be controlled by antivirals. In addition the DNA virus HPV causes tumors that depend on the continued expression of viral genes. Here, I will present data demonstrating that several of these viruses can be efficiently cleaved and destroyed using viral vectors that express Cas9 and virus-specific guide RNAs, thus providing a potential novel approach to treatment.

  • HBV – as target intervention
  • CRISPR/Cas9 RNA Guided Nuclease System (RGN)
  • NGG- PAM >> dsDNA cleavage >> NHEJ Repair >> Prfect repair >> Completion Repair Cycle
  • vs Mutagenesis – cycle exit
  1. HBV – Inactivation with CRISPR/Cas9: HBV lifecycle of the virus — reverse transcripatse (RT) of caspid and envelope – release antigens in blood invisible to Immune response, SUrfacce Antigen,COre Antigen, X-protein,
  2. Inhibitors of HBV  – cccDNA is stable for decades – virus in blood do not create new virus
  3. tetracycline represses HBV expression
  4. HBsAG va HBeAG: on core, surface, RT and N.S. sgRNA

Vector Delivery Strategies to the Liver

  • Strep pyogenes Cas9 – a diffrence PAM (5′ – NNGRRT-3′) – too large – ~4.8kb, including th ITRs.
  • Can we identify smaller Promoters
  • Packaging Sau Cas9 and two sgRNAs into AAV


  • using HBV-infected hepatocytes of a humanized liver or transgenic mouse – HBV Dual Target

Using HBV with RT and CRISPR Cas9 — viral load reduced to awake the immune response – allergy stage  – potential for future therapeutics



9:50 Grand Opening Coffee Break in the Exhibit Hall with Poster Viewing

10:35 Targeted Endonucleases as Antiviral Agents: Promises and Pitfalls

Keith R. Jerome, M.D., Ph.D., Member, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center; Professor and Head, Virology Division, Department of Laboratory Medicine, University of Washington

Genome editing offers the prospect of cure for infections such as HIV, hepatitis B virus, herpes simplex, and human papillomavirus, by disruption of essential viral nucleic acids or the human genes encoding receptors needed for viral entry. This talk will highlight the most recent laboratory data and the challenges still ahead in bringing this technology to the clinic.

  • Anti HSV – episonal DNA in Neuron, Acylovar – willingness to Participate in studies
  • Anti-HBV –
  • cART – HIV –

Viral replication returns if medication is taken away in two weeks

Herpes Simplex Virus HSV -1 – 50% HSV-2 16%

  1. CRISPR/Cas – difficult vectorization
  2. Targeted endonucleases
  3. Rare-cutting endonucleases in Gene Therapy – target distruction
  4. Derived from Crel enzyme by CELLECTIS AG (Paris) – co-expressed with Trex2 to remove 3″ hangover
  5. AAV as a targeted endonuclease delivery vector – mediated delivery to primary neuronal cultures
  6. Exposure to HSV-specific HE decreases virus production from neuronal cultures – virus production in treated cells
  • HSV-specific HE can disrupt HSV at all stages of the replication cycle – established in vivo
  1. cell planting
  2. AAV-mediated transgene delivery to the mouse TG in vivo – injection whiskerpad – eye scarified vs Eye not scarified
  3. AAV serotype: Transgene expression in TG is highly dependent on AAV serotype
  4. Dose dependence – Trigeminal ganglion (TG) – AAV-mediated transgene delivery to all branches ot TG
  5. In vivo mutugenesis of latent HSV
  6. Specificity – NGS analysis of Off target activity of NV1: Insertion vs Deletion vs NGS analysis of On and Off target activity of HSV1m8
  7. Endonuclease therapy suppresses viral reactivation
  8. Viral eradication: critical determinant: 3 of doses before cure occurs


Delivery system is the most important factor


11:05 Nucleic Acid Delivery Systems for RNA Therapy and Gene Editing

Daniel Anderson, Ph.D., Professor, Department of Chemical Engineering, Institute for Medical Engineering & Science, Harvard-MIT Division of Health Sciences & Technology and David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology

High throughput, combinatorial approaches have revolutionized small molecule drug discovery. Here we describe our high throughput methods for developing and characterizing RNA delivery and gene editing systems. Libraries of degradable polymers and lipid-like materials have been synthesized, formulated and screened for their ability to deliver RNA, both in vitro and in vivo. A number of delivery formulations have been developed with in vivo efficacy, and show potential applications for the treatment of genetic diseases, viral infections and cancers.

  1. Nanoparticulate approaches – repair your DNA while you still use it
  2. Barriers to Intracellular Delivery – What organs are most amenable to Targeting
  3. Liver, spleen, bone marrow, kidney
  4. Intracellular Drug Delivery:
  • Modular Pharmaci=ology with siRNA siRNA silences mRNA
  • Turning Nuclaic Acids into Drugs: Sequence Selection,  Mechanical modification (ligand conjugation), Encapsulation

Materials used for RNA Delivery – increase diversity of materials

  • Liquid light material: Combinatorial synthesis of lipid-like materials
  • RNA NAnoparticles – Lipid -siRNA-Nanoformulations targeting TTR in the liver of Primates
  • Mechanism of ApoE mediated iLNP Delivery [Phil Sharp/Alnylam]
  • si delivery to ENdothelium
  • Lipid modified Polymers: Short amino polymers – Total Dose 5 siRNA
  • Nanoformulation Chemistry: Endothelium in many organs(preferred) vs Hepatocytes
  • Immune cells as a target: CD45 or control
  • In vivo mediated Homologous Recombination Gene repair:  Nanoformulation deliver sgRNA
  1. In vivo delivery of sgRNA to endothelium wiht nanoparticle: mediated guide RNA delivery to endothilium: DNA Repair and Protein delivery
  2. Can CRISPR be used to repair a disease gene in vivo – PLASMID encoding Cas9 and GuideRNA + 199nt ss DNA repair template
  3. in vivo CRISPR rescue repairs defect, restores body weight and stops
  4. mRNA with nanoparticles: In vivo delivery- EPO Protein: Mean Human EPO
  • AAV Only
  • AAV +CAs9 – nano – 6% repain is therapeutic
  • In vivo mediated Gene Knockout without the Virus
  1. PCSK( Blood Cholesterol Liver PCSK9 Analysis – 60% gene mutated 65% reduction in CHolesterol — Synthetic system to do gene knockout


11:35 PANEL DISCUSSION: CRISPR/Cas: A Realistic and Practical Look at What the Future Could Hold

Moderator: Bryan R. Cullen, Ph.D., James B. Duke Professor of Molecular Genetics and Microbiology and Director, Center for Virology, Duke University

Participants: Session Speakers

Each speaker will spend a few minutes sharing their viewpoints and experiences on where things stand with using the CRISPR/Cas system for in vivo applications. Attendees will have an opportunity to ask questions and share their opinions.


  • Ex vivo delivery to Immune cell rather than to the tumor itself
  • solid cells therapy needs to reach each cell alternatives are needed

12:05 pm CRISPR/Cas9 for the Screening of the Human Kinome – A Pilot Study in an Aggressive Pediatric Cancer Cell Line

Simone T. Sredni, M.D., Ph.D., Research Associate Professor, Neurological Surgery, Northwestern University Feinberg School of Medicine, Ann and Robert H. Lurie Children’s Hospital of Chicago

The CRISPR-Cas9 system for genome editing is a powerful tool to identify genes involved in vital biological processes. A systematic functional screening of the human kinome has the potential to reveal molecules that are essential for tumor survival, growth, and migration. We will describe our experience using the Invitrogen LentiArray™CRISPR library to mutate 160kinases in a highly malignant pediatric tumor cell line. We will discuss our approach for screening, monitoring of cells lines, and validation.

  • LentiArray CRISPR Kinase Library – Bet Test 160 Kinase inhibitors
  • MRT – Malignant Rhabdoid Tumors – Children lexx 3 Years old
  • Genetic landmark, histology anatomical location: Kidney, Brain, Spine
  • Kinase Inhibitors and MRT
  • Finding Novel Targets
  1. Invitrogen – lentiArray CRISPR Library – edit 160 kinase genes – Viral Vector design
  • Cas9
  • gRNA-Kinase
  • Positive Control
  • Negative Control

2.  Kinome Screening – Impact on Proliferation 160 – only EIGHT were tested – significal=nly impaired cell profiferation

Retransaction and Confirmation of the identified  targets

PIM – Leukemia virus induce lymphomas: Proviral – PIM-1,2,3 KO

  • Verification of Genome Editing
  • PIMs – Proliferation
  • PIMs – Senescence – Adult hematological diseases and refractory solid tumors
  • PLK4 – Direct Mitosis Regulator – Activated Protein (mRNA) – expression in cytoplasm
  • PLK-4 and Cancer: Over-expression vs Deregulation – Colony Formation – colonygenic
  • Gene expression – Frozen Tumors – abnormality in children and in adults
  • Verification of Gene Editing: Cleavage and deletion
  • PLK-4 as a Cancer Drug Target – inhibitor enzymatic  – PLK-4 Inhibitor Xenografs





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Coronary Heart Disease Research: Sugar Industry influenced national conversation on heart disease – Adoption of Low Fat Diet vs Low Carbohydrates Diet

Reporter: Aviva Lev-Ari, PhD, RN

Public Health Outcome:

  • Uncontrolled consumption of sugar prevailed 1965 – 2005 – role of sugar in CVD was played down


  • Consumption of fat become the diet factor to be control and monitored in the Medical community – role of Fat was the main focus and its management by Statins


  • FDA Food Pyramid evolution

USDA Food Pyramid History

In January 1977, after listening to the testimony of Ancel Keys and other doctors and scientists intent on promoting the unsupported Dietary Fat-Heart hypothesis, the Committee published the “Dietary Goals for the United States” recommending that all Americans reduce their fat, saturated fat and cholesterol consumption, and increase their carbohydrate consumption to 55-60% of daily calories.


Concerns that were raised with the first dietary recommendations 30 y ago have yet to be adequately addressed. The initial Dietary Goals for Americans (1977) proposed increases in carbohydrate intake and decreases in fat, saturated fat, cholesterol, and salt consumption that are carried further in the 2010 Dietary Guidelines Advisory Committee (DGAC) Report. Important aspects of these recommendations remain unproven, yet a dietary shift in this direction has already taken place even as overweight/obesity and diabetes have increased. Although appealing to an evidence-based methodology, the DGAC Report demonstrates several critical weaknesses, including use of an incomplete body of relevant science; inaccurately representing, interpreting, or summarizing the literature; and drawing conclusions and/or making recommendations that do not reflect the limitations or controversies in the science. An objective assessment of evidence in the DGAC Report does not suggest a conclusive proscription against low-carbohydrate diets. The DGAC Report does not provide sufficient evidence to conclude that increases in whole grain and fiber and decreases in dietary saturated fat, salt, and animal protein will lead to positive health outcomes. Lack of supporting evidence limits the value of the proposed recommendations as guidance for consumers or as the basis for public health policy. It is time to reexamine how US dietary guidelines are created and ask whether the current process is still appropriate for our needs.



Curator: Aviva Lev-Ari, PhD, RN


UCSF reveals how sugar industry influenced national conversation on heart disease


Special Communication |

Sugar Industry and Coronary Heart Disease Research – A Historical Analysis of Internal Industry Documents

Cristin E. Kearns, DDS, MBA1,2; Laura A. Schmidt, PhD, MSW, MPH1,3,4; Stanton A. Glantz, PhD1,5,6,7,8
JAMA Intern Med. Published online September 12, 2016. doi:10.1001/jamainternmed.2016.5394

Corresponding Author: Stanton A. Glantz, PhD, UCSF Center for Tobacco Control Research and Education, 530 Parnassus Ave, Ste 366, San Francisco, CA 94143-1390 (glantz@medicine.ucsf.edu).

Accepted for Publication: July 2, 2016.

Published Online: September 12, 2016. doi:10.1001/jamainternmed.2016.5394

Author Contributions: Drs Kearns and Glantz had full access to all the data in the study and take responsibility for the integrity of the data and the accuracy of data analysis.

Early warning signals of the coronary heart disease (CHD) risk of sugar (sucrose) emerged in the 1950s. We examined Sugar Research Foundation (SRF) internal documents, historical reports, and statements relevant to early debates about the dietary causes of CHD and assembled findings chronologically into a narrative case study. The SRF sponsored its first CHD research project in 1965, a literature review published in the New England Journal of Medicine, which singled out fat and cholesterol as the dietary causes of CHD and downplayed evidence that sucrose consumption was also a risk factor. The SRF set the review’s objective, contributed articles for inclusion, and received drafts. The SRF’s funding and role was not disclosed. Together with other recent analyses of sugar industry documents, our findings suggest the industry sponsored a research program in the 1960s and 1970s that successfully cast doubt about the hazards of sucrose while promoting fat as the dietary culprit in CHD. Policymaking committees should consider giving less weight to food industry–funded studies and include mechanistic and animal studies as well as studies appraising the effect of added sugars on multiple CHD biomarkers and disease development.

These internal documents show that the SRF initiated CHD research in 1965 to protect market share and that its first project, a literature review, was published in NEJM in 1967 without disclosure of the sugar industry’s funding or role. The NEJM review served the sugar industry’s interests by arguing that epidemiologic, animal, and mechanistic studies associating sucrose with CHD were limited, implying they should not be included in an evidentiary assessment of the CHD risks of sucrose. Instead, the review argued that the only evidence modality needed to yield a definitive answer to the question of how to modify the American diet to prevent CHD was RCTs that exclusively used serum cholesterol level as a CHD biomarker. Randomized clinical trials using serum cholesterol level as the CHD biomarker made the high sucrose content of the American diet seem less hazardous than if the entire body of evidence had been considered.

Following the NEJM review, the sugar industry continued to fund research on CHD and other chronic diseases “as a main prop of the industry’s defense.”51 For example, in 1971, it influenced the National Institute of Dental Research’s National Caries Program to shift its emphasis to dental caries interventions other than restricting sucrose.8 The industry commissioned a review, “Sugar in the Diet of Man,” which it credited with, among other industry tactics, favorably influencing the 1976 US Food and Drug Administration evaluation of the safety of sugar.51 These findings, our analysis, and current Sugar Association criticisms of evidence linking sucrose to cardiovascular disease6,7 suggest the industry may have a long history of influencing federal policy.

This historical account of industry efforts demonstrates the importance of having reviews written by people without conflicts of interest and the need for financial disclosure. Scientific reviews shape policy debates, subsequent investigations, and the funding priorities of federal agencies.52 The NEJM has required authors to disclose all conflicts of interest since 1984,53 and conflict of interest disclosure policies have been widely implemented since the sugar industry launched its CHD research program. Whether current conflict of interest policies are adequate to withstand the economic interests of industry remains unclear.54

Many industries sponsor research to influence assessments of the risks and benefits of their products.55– 57The influence of industry sponsorship on nutrition research is receiving increased scrutiny.58 Access to documents not meant for public consumption has provided the public health community unprecedented insight into industry motives, strategies, tactics, and data designed to protect companies from litigation and regulation.59 This insight has been a major factor behind successful global tobacco control policies.60 Our analysis suggests that research using sugar industry documents has the potential to inform the health community about how to counter this industry’s strategies and tactics to control information on the adverse health effects of sucrose.

Study Limitations

The Roger Adams papers and other documents used in this research provide a narrow window into the activities of 1 sugar industry trade association; therefore, it is difficult to validate that the documents gathered are representative of the entirety of SRF internal materials related to Project 226 from the 1950s and 1960s or that the proper weight was given to each data source. There is no direct evidence that the sugar industry wrote or changed the NEJM review manuscript; the evidence that the industry shaped the review’s conclusions is circumstantial. We did not analyze the role of other organizations, nutrition leaders, or food industries that advocated that saturated fat and dietary cholesterol were the main dietary cause of CHD. We could not interview key actors involved in this historical episode because they have died.

This study suggests that the sugar industry sponsored its first CHD research project in 1965 to downplay early warning signals that sucrose consumption was a risk factor in CHD. As of 2016, sugar control policies are being promulgated in international,61 federal,62,63 state, and local venues.64 Yet CHD risk is inconsistently cited as a health consequence of added sugars consumption. Because CHD is the leading cause of death globally, the health community should ensure that CHD risk is evaluated in future risk assessments of added sugars. Policymaking committees should consider giving less weight to food industry–funded studies, and include mechanistic and animal studies as well as studies appraising the effect of added sugars on multiple CHD biomarkers and disease development.65



Council on Foods and Nutrition (American Medical Association).  The regulation of dietary fat: a report of the council. JAMA. 1962;181(5):411-429.
Link to Article

Yudkin  J. Pure, White and Deadly: The Problem of Sugar. London, England: Davis-Poynter Ltd; 1972.

Yudkin  J.  Diet and coronary thrombosis hypothesis and fact. Lancet. 1957;273(6987):155-162.
PubMed   |  Link to Article

Yudkin  J.  Dietary fat and dietary sugar in relation to ischaemic heart-disease and diabetes. Lancet. 1964;2(7349):4-5.
PubMed   |  Link to Article

Technical Group of Committee on Lipoproteins and Atherosclerosis and Committee on Lipoproteins and Atherosclerosis of National Advisory Heart Council.  Evaluation of serum lipoprotein and cholesterol measurements as predictors of clinical complications of atherosclerosis: report of a cooperative study of lipoproteins and atherosclerosisCirculation. 1956;14(4, pt 2):691-742.

Albrink  MJ.  Carbohydrate metabolism in cardiovascular disease. Ann Intern Med. 1965;62(6):1330-1333.
PubMed   |  Link to Article

Taubes  G, Couzens  CK. Big sugar’s sweet little lies: how the industry kept scientists from asking, does sugar kill? 2012. http://www.motherjones.com/environment/2012/10/sugar-industry-lies-campaign Accessed October 17, 2014.

Bero  L.  Implications of the tobacco industry documents for public health and policy. Annu Rev Public Health. 2003;24:267-288.
PubMed   |  Link to Article

US Department of Health and Human Services and US Department of Agriculture. 2015-2020 Dietary Guidelines for Americans. 8th ed. Washington, DC: U.S. Government Printing Office; 2016.

US Food and Drug Administration. Changes to the nutrition facts label. 2016.http://www.fda.gov/Food/GuidanceRegulation/GuidanceDocumentsRegulatoryInformation/LabelingNutrition/ucm385663.htm. Accessed June 7, 2016.

Miller  M, Stone  NJ, Ballantyne  C,  et al; American Heart Association Clinical Lipidology, Thrombosis, and Prevention Committee of the Council on Nutrition, Physical Activity, and Metabolism; Council on Arteriosclerosis, Thrombosis and Vascular Biology; Council on Cardiovascular Nursing; Council on the Kidney in Cardiovascular Disease.  Triglycerides and cardiovascular disease: a scientific statement from the American Heart AssociationCirculation. 2011;123(20):2292-2333.
PubMed   |  Link to Article

Teicholz  N. The Big Fat Surprise: Why Butter, Meat, and Cheese Belong in a Healthy Diet. New York, NY: Simon and Schuster; 2014.


Other related articles published in this Open Access Online Scientific Journal include the following:  


Metabolomics, Metabonomics and Functional Nutrition: The Next Step in Nutritional Metabolism and Biotherapeutics

Larry H. Bernstein, MD, FCAP  and Aviva Lev-Ari, PhD, RN

Reference Genes in the Human Gut Microbiome: The BGI Catalogue

Aviva Lev-Ari, PhD, RN

Two Mutations, in the PCSK9 Gene: Eliminates a Protein involved in Controlling LDL Cholesterol

Aviva Lev-Ari, PhD, RN

HDL-C: Target of Therapy – Steven E. Nissen, MD, MACC, Cleveland Clinic vs Peter Libby, MD, BWH

Aviva Lev-Ari, PhD, RN


The following articles in

Series A: e-Books on Cardiovascular Diseases

Series A Content Consultant: Justin D Pearlman, MD, PhD, FACC


Etiologies of Cardiovascular Diseases:

Epigenetics, Genetics and Genomics




Larry H Bernstein, MD, FCAP, Senior Editor, Author and Curator


Aviva Lev-Ari, PhD, RN, Editor and Curator


2.2.2: Endothelium, Angiogenesis, and Disordered Coagulation What is the Role of Plasma Viscosity in Hemostasis and Vascular Disease Risk? 

Larry H Bernstein, MD, FACP and Aviva Lev-Ari, PhD, RN Special Considerations in Blood Lipoproteins, Viscosity, Assessment and Treatment 

Larry H Bernstein, MD, FACP  and Aviva Lev-Ari, PhD, RN Biomarkers and risk factors for cardiovascular events, endothelial dysfunction, and thromboembolic complication

Larry H Bernstein, MD, FCAP A future for plasma metabolomics in cardiovascular disease assessment  

Larry H Bernstein, MD, FCAP Nitric Oxide Function in Coagulation – Part II

Larry H Bernstein, MD, FACP Nitric Oxide, Platelets, Endothelium and Hemostasis (Coagulation Part II)

Larry H Bernstein, MD, FACP Peroxisome Proliferator-Activated Receptor (PPAR-gamma) Receptors Activation: PPARγ Transrepression for Angiogenesis in Cardiovascular Disease and PPARγ Transactivation for Treatment of Diabetes 

Aviva Lev-Ari, PhD, RN

Endothelium Inflammatory Biomarkers Cardiovascular Risk: C-Reactive Protein BioMarker and Plasma Fibrinogen

Aviva Lev-Ari, PhD, RN Cardiovascular Risk Inflammatory Marker: Risk Assessment for Coronary Heart Disease and Ischemic Stroke ­ – Atherosclerosis

Aviva Lev-Ari, PhD, RN Importance of high sensitivity C-reactive protein (hs-CRP)

Larry H Bernstein, MD, FCAP


See also our Series A: Cardiovascular Diseases





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