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Archive for the ‘Genetics & Pharmaceutical’ Category


Reporter and Curator: Dr. Sudipta Saha, Ph.D.

 

Scientists think excessive population growth is a cause of scarcity and environmental degradation. A male pill could reduce the number of unintended pregnancies, which accounts for 40 percent of all pregnancies worldwide.

 

But, big drug companies long ago dropped out of the search for a male contraceptive pill which is able to chemically intercept millions of sperm before they reach a woman’s egg. Right now the chemical burden for contraception relies solely on the female. There’s not much activity in the male contraception field because an effective solution is available on the female side.

 

Presently, male contraception means a condom or a vasectomy. But researchers from Center for Drug Discovery at Baylor College of Medicine, USA are renewing the search for a better option—an easy-to-take pill that’s safe, fast-acting, and reversible.

 

The scientists began with lists of genes active in the testes for sperm production and motility and then created knockout mice that lack those genes. Using the gene-editing technology called CRISPR, in collaboration with Japanese scientists, they have so far made more than 75 of these “knockout” mice.

 

They allowed these mice to mate with normal (wild type) female mice, and if their female partners don’t get pregnant after three to six months, it means the gene might be a target for a contraceptive. Out of 2300 genes that are particularly active in the testes of mice, the researchers have identified 30 genes whose deletion makes the male infertile. Next the scientists are planning a novel screening approach to test whether any of about two billion chemicals can disable these genes in a test tube. Promising chemicals could then be fed to male mice to see if they cause infertility.

 

Female birth control pills use hormones to inhibit a woman’s ovaries from releasing eggs. But hormones have side effects like weight gain, mood changes, and headaches. A trial of one male contraceptive hormone was stopped early in 2011 after one participant committed suicide and others reported depression. Moreover, some drug candidates have made animals permanently sterile which is not the goal of the research. The challenge is to prevent sperm being made without permanently sterilizing the individual.

 

As a better way to test drugs, Scientists at University of Georgia, USA are investigating yet another high-tech approach. They are turning human skin cells into stem cells that look and act like the spermatogonial cells in the testes. Testing drugs on such cells might provide more accurate leads than tests on mice.

 

The male pill would also have to start working quickly, a lot sooner than the female pill, which takes about a week to function. Scientists from University of Dundee, U.K. admitted that there are lots of challenges. Because, a women’s ovary usually release one mature egg each month, while a man makes millions of sperm every day. So, the male pill has to be made 100 percent effective and act instantaneously.

 

References:

 

https://www.technologyreview.com/s/603676/the-search-for-a-perfect-male-birth-control-pill/

 

https://futurism.com/videos/the-perfect-male-birth-control-pill-is-coming-soon/?utm_source=Digest&utm_campaign=c42fc7b9b6-EMAIL_CAMPAIGN_2017_03_20&utm_medium=email&utm_term=0_03cd0a26cd-c42fc7b9b6-246845533

 

http://www.telegraph.co.uk/women/sex/the-male-pill-is-coming—and-its-going-to-change-everything/

 

http://www.mensfitness.com/women/sex-tips/male-birth-control-pill-making

 

http://health.howstuffworks.com/sexual-health/contraception/male-bc-pill.htm

 

http://europe.newsweek.com/male-contraception-side-effects-study-pill-injection-518237?rm=eu

 

http://edition.cnn.com/2016/01/07/health/male-birth-control-pill/index.html

 

http://www.nhs.uk/Conditions/contraception-guide/Pages/male-pill.aspx

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p53 mutation – Li-Fraumeni Syndrome – Likelihood of Genetic or Hereditary conditions playing a role in Intergenerational incidence of Cancer

 

Reporter: Aviva Lev-Ari, PhD, RN

 

THIS ARTICLE IS RECOMMENDED READING TO ALL OUR e-Readers

because it is a REAL story of a high school student fighting Brain Cancer, glioblastoma multiforme (GBM)

it presents the FRONTIER OF GENOMICS, PRECISION MEDICINE, Interventional Radiology and Interventional ONCOLOGY at

Stanford University, Canary Center at Stanford for Early Cancer Detection, Stanford Medical Center and Lucile Packard Children’s Hospital

I was exposed to Li-Fraumeni Syndrome in the following article:

‘And yet, you try’ – A father’s quest to save his son

http://stanmed.stanford.edu/2016fall/milan-gambhirs-li-fraumeni-syndrome.html

 

Li-Fraumeni syndrome

Other Names for This Condition

  • LFS
  • Sarcoma family syndrome of Li and Fraumeni
  • Sarcoma, breast, leukemia, and adrenal gland (SBLA) syndrome
  • SBLA syndrome

LFS is a rare disorder that greatly increases the risk of developing several types of cancer, particularly in children and young adults.

The cancers most often associated with Li-Fraumeni syndrome include breast cancer, a form of bone cancer called osteosarcoma, and cancers of soft tissues (such as muscle) called

Soft tissue sarcoma forms in soft tissues of the body, including muscle, tendons, fat, blood vessels, lymph vessels, nerves, and tissue around joints.


(small hormone-producing glands on top of each kidney). Several other types of cancer also occur more frequently in people with Li-Fraumeni syndrome.

A very similar condition called Li-Fraumeni-like syndrome shares many of the features of classic Li-Fraumeni syndrome. Both conditions significantly increase the chances of developing multiple cancers beginning in childhood; however, the pattern of specific cancers seen in affected family members is different.

Genetic Changes

The CHEK2 and TP53 genes are associated with Li-Fraumeni syndrome.

More than half of all families with Li-Fraumeni syndrome have inherited mutations in the gene. TP53 is a tumor suppressor gene, which means that it normally helps control the growth and division of cells. Mutations in this gene can allow cells to divide in an uncontrolled way and form tumors. Other genetic and environmental factors are also likely to affect the risk of cancer in people with TP53 mutations.

A few families with cancers characteristic of Li-Fraumeni syndrome and Li-Fraumeni-like syndrome do not have TP53 mutations, but have mutations in the CHEK2 gene. Like the TP53 gene, CHEK2 is a tumor suppressor gene. Researchers are uncertain whether CHEK2 mutations actually cause these conditions or are merely associated with an increased risk of certain cancers (including breast cancer).

Inheritance Pattern

Li-Fraumeni syndrome is inherited in an autosomal dominant pattern, which means one copy of the altered gene in each cell is sufficient to increase the risk of developing cancer. In most cases, an affected person has a parent and other family members with cancers characteristic of the condition.

Diagnosis and Management

These resources address the diagnosis or management of Li-Fraumeni syndrome:

References on LFS

SOURCE

https://ghr.nlm.nih.gov/condition/li-fraumeni-syndrome

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Reporter and Curator: Dr. Sudipta Saha, Ph.D.

 

MicroRNAs (miRNAs) are a group of small non-coding RNA molecules that play a major role in posttranscriptional regulation of gene expression and are expressed in an organ-specific manner. One miRNA can potentially regulate the expression of several genes, depending on cell type and differentiation stage. They control every cellular process and their altered regulation is involved in human diseases. miRNAs are differentially expressed in the male and female gonads and have an organ-specific reproductive function. Exerting their affect through germ cells and gonadal somatic cells, miRNAs regulate key proteins necessary for gonad development. The role of miRNAs in the testes is only starting to emerge though they have been shown to be required for adequate spermatogenesis. In the ovary, miRNAs play a fundamental role in follicles’ assembly, growth, differentiation, and ovulation.

 

Deciphering the underlying causes of idiopathic male infertility is one of the main challenges in reproductive medicine. This is especially relevant in infertile patients displaying normal seminal parameters and no urogenital or genetic abnormalities. In these cases, the search for additional sperm biomarkers is of high interest. This study was aimed to determine the implications of the sperm miRNA expression profiles in the reproductive capacity of normozoospermic infertile individuals. The expression levels of 736 miRNAs were evaluated in spermatozoa from normozoospermic infertile males and normozoospermic fertile males analyzed under the same conditions. 57 miRNAs were differentially expressed between populations; 20 of them was regulated by a host gene promoter that in three cases comprised genes involved in fertility. The predicted targets of the differentially expressed miRNAs unveiled a significant enrichment of biological processes related to embryonic morphogenesis and chromatin modification. Normozoospermic infertile individuals exhibit a specific sperm miRNA expression profile clearly differentiated from normozoospermic fertile individuals. This miRNA cargo has potential implications in the individuals’ reproductive competence.

 

Circulating or “extracellular” miRNAs detected in biological fluids, could be used as potential diagnostic and prognostic biomarkers of several disease, such as cancer, gynecological and pregnancy disorders. However, their contributions in female infertility and in vitro fertilization (IVF) remain unknown. Polycystic ovary syndrome (PCOS) is a frequent endocrine disorder in women. PCOS is associated with altered features of androgen metabolism, increased insulin resistance and impaired fertility. Furthermore, PCOS, being a syndrome diagnosis, is heterogeneous and characterized by polycystic ovaries, chronic anovulation and evidence of hyperandrogenism, as well as being associated with chronic low-grade inflammation and an increased life time risk of type 2 diabetes. Altered miRNA levels have been associated with diabetes, insulin resistance, inflammation and various cancers. Studies have shown that circulating miRNAs are present in whole blood, serum, plasma and the follicular fluid of PCOS patients and that these might serve as potential biomarkers and a new approach for the diagnosis of PCOS. Presence of miRNA in mammalian follicular fluid has been demonstrated to be enclosed within microvesicles and exosomes or they can also be associated to protein complexes. The presence of microvesicles and exosomes carrying microRNAs in follicular fluid could represent an alternative mechanism of autocrine and paracrine communication inside the ovarian follicle. The investigation of the expression profiles of five circulating miRNAs (let-7b, miR-29a, miR-30a, miR-140 and miR-320a) in human follicular fluid from women with normal ovarian reserve and with polycystic ovary syndrome (PCOS) and their ability to predict IVF outcomes showed that these miRNAs could provide new helpful biomarkers to facilitate personalized medical care for oocyte quality in ART (Assisted Reproductive Treatment) and during IVF (In Vitro Fertilization).

 

References:

 

http://link.springer.com/chapter/10.1007%2F978-3-319-31973-5_12

 

http://onlinelibrary.wiley.com/doi/10.1111/andr.12276/abstract;jsessionid=F805A89DCC94BDBD42D6D60C40AD4AB0.f03t03

 

http://www.sciencedirect.com/science/article/pii/S0009279716302241

 

http://link.springer.com/article/10.1007%2Fs10815-016-0657-9

 

http://www.nature.com/articles/srep24976

 

 

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Real Time Coverage and eProceedings of Presentations on 9/19-9/21 @CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

Curator: Aviva Lev-Ari, PhD, RN

 

LIVE 9/19 8AM – 10AM USING CRISPR/Cas9 FOR FUNCTIONAL SCREENING at CHI’s 2nd Annual Symposium CRISPR: Mechanisms and Applications @CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

https://pharmaceuticalintelligence.com/2016/09/19/live-919-8am-10am-using-crisprcas9-for-functional-screening-at-chis-2nd-annual-symposium-crispr-mechanisms-and-applications-chis-14th-discovery-on-target-919-9222/

 

LIVE 9/19 9:40 – noon CRISPR Engineering Lymphoma Lines & Will Interference from CRISPR Silence RNAi? CHI’s 2nd Annual Symposium CRISPR: Mechanisms and Applications @ CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

https://pharmaceuticalintelligence.com/2016/09/19/live-919-940-noon-crispr-engineering-lymphoma-lines-will-interference-from-crispr-silence-rnai-chis-2nd-annual-symposium-crispr-mechanisms-and-applications-chis-14th/

 

LIVE 9/19 1:40 – 3:20 EMERGING APPLICATIONS OF CRISPR/CAS9 at CHI’s 2nd Annual Symposium CRISPR: Mechanisms and Applications @ CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

https://pharmaceuticalintelligence.com/2016/09/19/live-919-140-320-emerging-applications-of-crisprcas9-at-chis-2nd-annual-symposium-crispr-mechanisms-and-applications-chis-14th-discovery-on-target-919-9222016/

 

LIVE 9/19 4PM – 5:30PM NK CELL-BASED CANCER IMMUNOTHERAPY @CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

https://pharmaceuticalintelligence.com/2016/09/19/live-919-4pm-530pm-nk-cell-based-cancer-immunotherapy-chis-14th-discovery-on-target-919-9222016-westin-boston-waterfront-boston/

 

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

https://pharmaceuticalintelligence.com/2016/09/20/live-920-8am-to-noon-gene-therapies-breakthroughs-at-chis-14th-discovery-on-target-919-9222016-westin-boston-waterfront-boston/

 

LIVE 9/20 2PM to 5:30PM New Viruses for Therapeutic Gene Delivery at CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

https://pharmaceuticalintelligence.com/2016/09/20/live-920-2pm-to-530pm-new-viruses-for-therapeutic-gene-delivery-at-chis-14th-discovery-on-target-919-9222016-westin-boston-waterfront-boston/

 

LIVE 9/21 8AM to 10:55 AM Expoloring the Versatility of CRISPR/Cas9 at CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

https://pharmaceuticalintelligence.com/2016/09/21/live-921-8am-to-1055-am-expoloring-the-versatility-of-crisprcas9-at-chis-14th-discovery-on-target-919-9222016-westin-boston-waterfront-boston/

 

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

https://pharmaceuticalintelligence.com/2016/09/21/live-921-8am-to-240pm-targeting-cardio-metabolic-diseases-a-focus-on-liver-fibrosis-and-nash-targets-at-chis-14th-discovery-on-target-919-9222016-westin-boston-waterfront-b/

 

LIVE 9/21 12:50 pm Plenary Keynote Program at CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

https://pharmaceuticalintelligence.com/2016/09/21/live-921-1250-pm-plenary-keynote-program-at-chis-14th-discovery-on-target-919-9222016-westin-boston-waterfront-boston/

 

LIVE 9/21 3:20PM to 6:40PM KINASE INHIBITORS FOR CANCER IMMUNOTHERAPY COMBINATIONS & KINASE INHIBITORS FOR AUTOIMMUNE AND INFLAMMATORY DISEASES at CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

https://pharmaceuticalintelligence.com/2016/09/21/live-921-320pm-to-640pm-kinase-inhibitors-for-cancer-immunotherapy-combinations-kinase-inhibitors-for-autoimmune-and-inflammatory-diseases-at-chis-14th-discovery-on-target-919/

 

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LIVE 9/20 2PM to 5:30PM New Viruses for Therapeutic Gene Delivery at CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

http://www.discoveryontarget.com/

http://www.discoveryontarget.com/crispr-therapies/

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

Editor-in-Chief

http://pharmaceuticalintelligence.com

#BostonDOT16

@BostonDOT

 

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

 

ALS  

  • 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

www.pactgroup.net

 

They do a standard panel of tests for viral infections.

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

 

@BeckmanInst

@cityofhope

 

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

12:45 PM Screening with shRNA and CRISPR

Ryan Raver, PhD Global Product Manager, Functional Genomics, MilliporeSigma

  • KO – Knock Out
  • KD – Knock Down
  1. RNAi -KD
  2. CRISPR-Cas9 – KO

NEW STRATEGIES FOR BETTER SPECIFICITY AND DELIVERY

2:05 Chairperson’s Remarks

Joseph Gold, Center for Biomedicine and Genetics Beckman Research Institute, City of Hope

2:15 Large Scale Production of cell Therapies for Regenerative Medicine: COmbination Cell and Gene Therapy products

Joseph Gold, Center for Biomedicine and Genetics Beckman Research Institute, City of Hope

  • Biological & Cellular GMP manufacturing Core at COH
  • Establishing scalable hESC suspension Culture
  • Optimized small molecule concentration, induction timing, stirring rates
  1. Almost Xeno free
  2. defined
  3. very good reproducibility
  4. high purity and yield:
  • Immuno-staining,
  • FACS – cTnT, sMHC, Alpha-actinin
  • Cryopreservation, Multi-electrode Array (MEA)
  • hESC-RPE monolayer on synthetic substrate

Combination cell/gene therapy products at COH

  1. CAR T CCR5-inactivated CD34+ HSPC – Target: AIDS
  • adoptive immunotherapy using CAR-Engineering T cells – glioblastoma

2. HIV resistance with engineering CCR5-negative blood stem cells: gene KO by ZFNs

  • assay considerations 0 If cryopreservation : Identify, viral insertion, endotoxin, residual beads Potency: CAR T- % transduced cells, CCR5-?-CD34 cells: HIV resistance

3. Glioblastoma

4.  In vivo activity of transduced NSCs: Assay consideration – viability, sterility, mycobatom,

5.  ALS: degeneration of neurones

  • Embryonic stem cells
  • Fetal neural stem cells
  • Adult stem cells – human Proginetor neural cells

6.  Canavan Disease – Y.Shi – ASPA gene mutations cause Canavan disease

Strategy: autologous iPSC-derived, modified neural progenitors: DIferentiate to neural progenitors

Gene therapy: Correction (Off-taregt effects, correct individual cell lines)  or Over expression (Copy number, selection, transduce iPSC)

  • release assay consideration
  • identity – markers and HLA, contaminants, Potency: in vivo efficacy modified autologous
  • production Assistance for Cell therapy
  • cell therapy manufacturing development
  • roles of Biobanks

 

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; Director, Berkeley Stem Cell Center, University of California, Berkeley

Adeno-associated viral (AAV) vectors have been increasingly successful in clinical trials; however, viruses face many delivery barriers that limit their efficacy for most disease targets. We have developed directed vector evolution – the iterative genetic diversification of a viral genome and functional selection for desired properties – to engineer novel, optimized AAV vectors for efficient, selective delivery for a range of tissue and disease targets.

  • DNA (gene therapy and editing) >> RNAi (antisense) >> Protein: Small molecules and monoclonal antibodies
  • Dlivery
  • Adeno Associated Viral Vectors Adenoviral helper genes
  • AAV2, efficacy in Leber’s Congenital samaurosis (AAV)
  • Spinal muscular, lipolrotein lipase deficiency (AAV)

Gene Delivery

  1. neutralization of pre-esisting antibodies
  2. target tissue – deep penetration
  3. Inefficient transduction to target cells
  4. target specific cells

Fitness as Therapy – virus evolutionary: Tropism and immunity

  • AAV directed Vector evolution : Input /sequence >> Diversity/generation >> Packaging
  • Retinoschisis Model – Eye therapeutic gene to protect vision – AAV – transduction of retinal cell, vitreas in Human different scale — eventually Macualr degeneration Therapy for: Photo receptor is the target for therapy
  • Dog: Fundus Imaging of Engineered AAV: Variant Expression GFP Pool Carrying

Lung- Cystic Fibrosis (mucus production amplifies (enhanced transduction) due to MAC3 translation error) – Gene Therapy – cilia function to restore ability to clear mucos: Variant evolved on Human airway epithelia – AA  particles

  1. AAV2>>> AAV%>> T mutation
  • efficacy must be improved

Brain and Spinal Cord

  1. Scull – 8 drills followed by 16 AAV injections
  2. Spinal cord: injuction in muscle

Synthetic version of AAV — Engineered AAV for enhanced Retrograde Transport

  • AAV2-retrograded transport – Noval variant Undergoes Transport along multiple Projections
  • Cas9 – Retrograde Delivery of Cas9 to cortex of mouse – KD –

Summary

  • Virus as gene delivery mechanism
  • designer AAV variants

 

3:15 Sponsored Presentation (Opportunity Available)

3:45 Refreshment Break in the Exhibit Hall with Poster Viewing and Poster Competition Winner Announced

4:25 Lentiviral Vectors for Gene Therapy

Manjunath N. Swamy, MD, Professor of Biomedical Sciences and Co-DIrector of the Center of Emphasis in Infectious Diseases. Paul L Foster School of Medicine, Texas Tech University Health Science Center

  • RNAi targets for HIV
  • Expression of multiple shRNAs
  • COnstruction od 7 shRNA-miR targeting CCR5 and 6 viral genes – protect against both 5 ans x4 tropic HIV-1
  • shRNA expression does not decrease with distance from promoter
  • Lentiviral vector to express ZFNs: HIV envelop – ZFN-mediated CCR5 gene editing in Primary T cells
  • ZFN treatment of HIV+PBMC prevents activation of HIV
  • Strategy to encapsulate Cas9 Protein wihtin LV : AIm to deliver Cas9 protein deliver sgRNA expression vector. A Lentiviral
  • Gene editing by all-in-one Lentivirus = to prepack

 

4:55 AAV Caspid Engineering

Miguel Sena Esteves, Ph.D., Associate Professor, Department of Neurology, Gene Therapy Center, University of Massachusetts Medical School

Adeno-associated virus vectors have become the leading platform for development of in vivo gene therapies for neurological diseases. We have developed new AAV vectors for widespread gene delivery to the CNS through vascular infusion in adult animals through peptide grafting and in vivo library selection. These new neurotropic AAVs have achieved CNS-wide silencing of gene expression using gene-specific microRNAs.

  • Adeno-associated virus – Paravovirus family
  • Recombinant AAV vectors carry gene expression cassette of choice flanked by two
  • CNS – route of gene delivery
  • Crossing BBB
  • Systemic delivery of AAV9 vectors: IV
  • Peptide grafting – in vivo selection of novel CNS: DIstribution of GFP transduced cells – robust neuronal transducture with transduction AAV-AS
  • motor cortex, Straiatum, thalamus, motor cortex, ventral horn of spinal cord, cerebelum, liver, muscle, oculomotor nerve, nucleus of oculomotor nerve

Huntington’s Disease > 40 CAG vs <26 CAG in normal – Peptide grafting of AAV vectors for CNS

  • DNA shuffling and in vivo selection: Brain vs Liver

Next generation of AAV vectors for CNS 

  • Liver, pancreas, lung — same pattern

neural transduction after vascular delivery

  • AAV-B1 caspid vs AAV8 – 19 amino acids

Conclusion

New capsids with improved CNS tropism

19 alanines modifies the CNS tropism of AAV9 variants

Chimeric caspids identified from in vivo screens

 

5:25 Welcome Reception in the Exhibit Hall with Poster Viewing

 

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Milestones in Physiology & Discoveries in Medicine and Genomics: Request for Book Review Writing on Amazon.com


physiology-cover-seriese-vol-3individualsaddlebrown-page2

Milestones in Physiology

Discoveries in Medicine, Genomics and Therapeutics

Patient-centric Perspective 

http://www.amazon.com/dp/B019VH97LU 

2015

 

 

Author, Curator and Editor

Larry H Bernstein, MD, FCAP

Chief Scientific Officer

Leaders in Pharmaceutical Business Intelligence

Larry.bernstein@gmail.com

Preface

Introduction 

Chapter 1: Evolution of the Foundation for Diagnostics and Pharmaceuticals Industries

1.1  Outline of Medical Discoveries between 1880 and 1980

1.2 The History of Infectious Diseases and Epidemiology in the late 19th and 20th Century

1.3 The Classification of Microbiota

1.4 Selected Contributions to Chemistry from 1880 to 1980

1.5 The Evolution of Clinical Chemistry in the 20th Century

1.6 Milestones in the Evolution of Diagnostics in the US HealthCare System: 1920s to Pre-Genomics

 

Chapter 2. The search for the evolution of function of proteins, enzymes and metal catalysts in life processes

2.1 The life and work of Allan Wilson
2.2  The  evolution of myoglobin and hemoglobin
2.3  More complexity in proteins evolution
2.4  Life on earth is traced to oxygen binding
2.5  The colors of life function
2.6  The colors of respiration and electron transport
2.7  Highlights of a green evolution

 

Chapter 3. Evolution of New Relationships in Neuroendocrine States
3.1 Pituitary endocrine axis
3.2 Thyroid function
3.3 Sex hormones
3.4 Adrenal Cortex
3.5 Pancreatic Islets
3.6 Parathyroids
3.7 Gastointestinal hormones
3.8 Endocrine action on midbrain
3.9 Neural activity regulating endocrine response

3.10 Genomic Promise for Neurodegenerative Diseases, Dementias, Autism Spectrum, Schizophrenia, and Serious Depression

 

Chapter 4.  Problems of the Circulation, Altitude, and Immunity

4.1 Innervation of Heart and Heart Rate
4.2 Action of hormones on the circulation
4.3 Allogeneic Transfusion Reactions
4.4 Graft-versus Host reaction
4.5 Unique problems of perinatal period
4.6. High altitude sickness
4.7 Deep water adaptation
4.8 Heart-Lung-and Kidney
4.9 Acute Lung Injury

4.10 Reconstruction of Life Processes requires both Genomics and Metabolomics to explain Phenotypes and Phylogenetics

 

Chapter 5. Problems of Diets and Lifestyle Changes

5.1 Anorexia nervosa
5.2 Voluntary and Involuntary S-insufficiency
5.3 Diarrheas – bacterial and nonbacterial
5.4 Gluten-free diets
5.5 Diet and cholesterol
5.6 Diet and Type 2 diabetes mellitus
5.7 Diet and exercise
5.8 Anxiety and quality of Life
5.9 Nutritional Supplements

 

Chapter 6. Advances in Genomics, Therapeutics and Pharmacogenomics

6.1 Natural Products Chemistry

6.2 The Challenge of Antimicrobial Resistance

6.3 Viruses, Vaccines and immunotherapy

6.4 Genomics and Metabolomics Advances in Cancer

6.5 Proteomics – Protein Interaction

6.6 Pharmacogenomics

6.7 Biomarker Guided Therapy

6.8 The Emergence of a Pharmaceutical Industry in the 20th Century: Diagnostics Industry and Drug Development in the Genomics Era: Mid 80s to Present

6.09 The Union of Biomarkers and Drug Development

6.10 Proteomics and Biomarker Discovery

6.11 Epigenomics and Companion Diagnostics

 

Chapter  7

Integration of Physiology, Genomics and Pharmacotherapy

7.1 Richard Lifton, MD, PhD of Yale University and Howard Hughes Medical Institute: Recipient of 2014 Breakthrough Prizes Awarded in Life Sciences for the Discovery of Genes and Biochemical Mechanisms that cause Hypertension

7.2 Calcium Cycling (ATPase Pump) in Cardiac Gene Therapy: Inhalable Gene Therapy for Pulmonary Arterial Hypertension and Percutaneous Intra-coronary Artery Infusion for Heart Failure: Contributions by Roger J. Hajjar, MD

7.3 Diagnostics and Biomarkers: Novel Genomics Industry Trends vs Present Market Conditions and Historical Scientific Leaders Memoirs

7.4 Synthetic Biology: On Advanced Genome Interpretation for Gene Variants and Pathways: What is the Genetic Base of Atherosclerosis and Loss of Arterial Elasticity with Aging

7.5 Diagnosing Diseases & Gene Therapy: Precision Genome Editing and Cost-effective microRNA Profiling

7.6 Imaging Biomarker for Arterial Stiffness: Pathways in Pharmacotherapy for Hypertension and Hypercholesterolemia Management

7.7 Neuroprotective Therapies: Pharmacogenomics vs Psychotropic drugs and Cholinesterase Inhibitors

7.8 Metabolite Identification Combining Genetic and Metabolic Information: Genetic association links unknown metabolites to functionally related genes

7.9 Preserved vs Reduced Ejection Fraction: Available and Needed Therapies

7.10 Biosimilars: Intellectual Property Creation and Protection by Pioneer and by

7.11 Demonstrate Biosimilarity: New FDA Biosimilar Guidelines

 

Chapter 7.  Biopharma Today

8.1 A Great University engaged in Drug Discovery: University of Pittsburgh

8.2 Introduction – The Evolution of Cancer Therapy and Cancer Research: How We Got Here?

8.3 Predicting Tumor Response, Progression, and Time to Recurrence

8.4 Targeting Untargetable Proto-Oncogenes

8.5 Innovation: Drug Discovery, Medical Devices and Digital Health

8.6 Cardiotoxicity and Cardiomyopathy Related to Drugs Adverse Effects

8.7 Nanotechnology and Ocular Drug Delivery: Part I

8.8 Transdermal drug delivery (TDD) system and nanotechnology: Part II

8.9 The Delicate Connection: IDO (Indolamine 2, 3 dehydrogenase) and Cancer Immunology

8.10 Natural Drug Target Discovery and Translational Medicine in Human Microbiome

8.11 From Genomics of Microorganisms to Translational Medicine

8.12 Confined Indolamine 2, 3 dioxygenase (IDO) Controls the Homeostasis of Immune Responses for Good and Bad

 

Chapter 9. BioPharma – Future Trends

9.1 Artificial Intelligence Versus the Scientist: Who Will Win?

9.2 The Vibrant Philly Biotech Scene: Focus on KannaLife Sciences and the Discipline and Potential of Pharmacognosy

9.3 The Vibrant Philly Biotech Scene: Focus on Computer-Aided Drug Design and Gfree Bio, LLC

9.4 Heroes in Medical Research: The Postdoctoral Fellow

9.5 NIH Considers Guidelines for CAR-T therapy: Report from Recombinant DNA Advisory Committee

9.6 1st Pitch Life Science- Philadelphia- What VCs Really Think of your Pitch

9.7 Multiple Lung Cancer Genomic Projects Suggest New Targets, Research Directions for Non-Small Cell Lung Cancer

9.8 Heroes in Medical Research: Green Fluorescent Protein and the Rough Road in Science

9.9 Issues in Personalized Medicine in Cancer: Intratumor Heterogeneity and Branched Evolution Revealed by Multiregion Sequencing

9.10 The SCID Pig II: Researchers Develop Another SCID Pig, And Another Great Model For Cancer Research

Epilogue

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metabolomics-seriesdindividualred-page2

Metabolic Genomics & Pharmaceutics

2015

http://www.amazon.com/dp/B012BB0ZF0

 

Author, Curator and Editor

Larry H Bernstein, MD, FCAP

Chief Scientific Officer

Leaders in Pharmaceutical Business Intelligence

Larry.bernstein@gmail.com

Chapter 1: Metabolic Pathways

1.1            Carbohydrate Metabolism

1.2            Studies of Respiration Lead to Acetyl CoA

1.3            Pentose Shunt, Electron Transfer, Galactose, more Lipids in brief

1.4            The Multi-step Transfer of Phosphate Bond and Hydrogen Exchange Energy

1.5            Diabetes Mellitus

1.6            Glycosaminoglycans, Mucopolysaccharides, L-iduronidase, Enzyme Therapy

Chapter 2: Lipid Metabolism

2.1            Lipid Classification System

2.2            Essential Fatty Acids

2.3            Lipid Oxidation and Synthesis of Fatty Acids

2.4            Cholesterol and Regulation of Liver Synthetic Pathways

2.5            Sex hormones, Adrenal cortisol, Prostaglandins

2.6            Cytoskeleton and Cell Membrane Physiology

2.7            Pharmacological Action of Steroid hormone

Chapter 3: Cell Signaling

3.1            Signaling and Signaling Pathways

3.2            Signaling Transduction Tutorial

3.3            Selected References to Signaling and Metabolic Pathways in Leaders in Pharmaceutical Intelligence

3.4            Integrins, Cadherins, Signaling and the Cytoskeleton

3.5            Complex Models of Signaling: Therapeutic Implications

3.6            Functional Correlates of Signaling Pathways

Chapter 4: Protein Synthesis and Degradation

4.1            The Role and Importance of Transcription Factors

4.2            RNA and the Transcription of the Genetic Code

4.3            9:30 – 10:00, 6/13/2014, David Bartel “MicroRNAs, Poly(A) tails and Post-transcriptional Gene Regulation

4.4            Transcriptional Silencing and Longevity Protein Sir2

4.5            Ca2+ Signaling: Transcriptional Control

4.6            Long Noncoding RNA Network regulates PTEN Transcription

4.7            Zinc-Finger Nucleases (ZFNs) and Transcription Activator–Like Effector Nucleases (TALENs)

4.8            Cardiac Ca2+ Signaling: Transcriptional Control

4.9            Transcription Factor Lyl-1 Critical in Producing Early T-Cell Progenitors

4.10            Human Frontal Lobe Brain: Specific Transcriptional Networks

4.11            Somatic, Germ-cell, and Whole Sequence DNA in Cell Lineage and Disease

Chapter 5:  Sub-cellular Structure

5.1            Mitochondria: Origin from Oxygen free environment, Role in Aerobic Glycolysis and Metabolic Adaptation

5.2            Mitochondrial Metabolism and Cardiac Function

5.3            Mitochondria: More than just the “Powerhouse of the Cell”

5.4            Mitochondrial Fission and Fusion: Potential Therapeutic Targets?

5.5            Mitochondrial Mutation Analysis might be “1-step” Away

5.6            Autophagy-Modulating Proteins and Small Molecules Candidate Targets for Cancer Therapy: Commentary of Bioinformatics Approaches

5.7            Chromatophagy, A New Cancer Therapy: Starve The Diseased Cell Until It Eats Its Own DNA

5.8           A Curated Census of Autophagy-Modulating Proteins and Small Molecules Candidate Targets for Cancer Therapy

5.9           Role of Calcium, the Actin Skeleton, and Lipid Structures in Signaling and Cell Motility

Chapter 6: Proteomics

6.1            Proteomics, Metabolomics, Signaling Pathways, and Cell Regulation: a Compilation of Articles in the Journal http://pharmaceuticalintelligence.com

6.2            A Brief Curation of Proteomics, Metabolomics, and Metabolism

6.3            Using RNA-seq and Targeted Nucleases to Identify Mechanisms of Drug Resistance in Acute Myeloid Leukemia, SK Rathe in Nature, 2014

6.4            Proteomics – The Pathway to Understanding and Decision-making in Medicine

6.5            Advances in Separations Technology for the “OMICs” and Clarification of Therapeutic Targets

6.6           Expanding the Genetic Alphabet and Linking the Genome to the Metabolome

6.7            Genomics, Proteomics and Standards

6.8            Proteins and Cellular Adaptation to Stress

6.9            Genes, Proteomes, and their Interaction

6.10           Regulation of Somatic Stem Cell Function

6.11           Scientists discover that Pluripotency factor NANOG is also active in Adult Organism

Chapter 7: Metabolomics

7.1            Extracellular Evaluation of Intracellular Flux in Yeast Cells

7.2            Metabolomic Analysis of Two Leukemia Cell Lines Part I

7.3            Metabolomic Analysis of Two Leukemia Cell Lines Part II

7.4            Buffering of Genetic Modules involved in Tricarboxylic Acid Cycle Metabolism provides Homeomeostatic Regulation

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

7.6            Isoenzymes in Cell Metabolic Pathways

7.7            A Brief Curation of Proteomics, Metabolomics, and Metabolism

7.8            Metabolomics is about Metabolic Systems Integration

7.9             Mechanisms of Drug Resistance

7.10           Development Of Super-Resolved Fluorescence Microscopy

7.11            Metabolic Reactions Need Just Enough

Chapter 8.  Impairments in Pathological States: Endocrine Disorders; Stress Hypermetabolism and CAncer

8.1           Omega3 Fatty Acids, Depleting the Source, and Protein Insufficiency in Renal Disease

8.2             Liver Endoplasmic Reticulum Stress and Hepatosteatosis

8.3            How Methionine Imbalance with Sulfur Insufficiency Leads to Hyperhomocysteinemia

8.4            AMPK Is a Negative Regulator of the Warburg Effect and Suppresses Tumor Growth InVivo

8.5           A Second Look at the Transthyretin Nutrition Inflammatory Conundrum

8.6            Mitochondrial Damage and Repair under Oxidative Stress

8.7            Metformin, Thyroid Pituitary Axis, Diabetes Mellitus, and Metabolism

8.8            Is the Warburg Effect the Cause or the Effect of Cancer: A 21st Century View?

8.9            Social Behavior Traits Embedded in Gene Expression

8.10          A Future for Plasma Metabolomics in Cardiovascular Disease Assessment

Chapter 9: Genomic Expression in Health and Disease 

9.1            Genetics of Conduction Disease: Atrioventricular (AV) Conduction Disease (block): Gene Mutations – Transcription, Excitability, and Energy Homeostasis

9.2            BRCA1 a Tumour Suppressor in Breast and Ovarian Cancer – Functions in Transcription, Ubiquitination and DNA Repair

9.3            Metabolic Drivers in Aggressive Brain Tumors

9.4            Modified Yeast Produces a Range of Opiates for the First time

9.5            Parasitic Plant Strangleweed Injects Host With Over 9,000 RNA Transcripts

9.6            Plant-based Nutrition, Neutraceuticals and Alternative Medicine: Article Compilation the Journal

9.7            Reference Genes in the Human Gut Microbiome: The BGI Catalogue

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

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

Summary 

Epilogue


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