Archive for the ‘Cell Processing System in Cell Therapy Process Development’ Category

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


During menopause a woman’s ovaries stop working—leading to hot flashes, sleep problems, weight gain, and worse, bone deterioration. Now scientists are exploring whether transplanting lab-made ovaries might stop those symptoms. In one of the first efforts to explore the potential of such a technique, researchers say they used tissue engineering to construct artificial rat ovaries able to supply female hormones like estrogen and progesterone. A research carried out at Wake Forest Baptist Medical Center, suggests a potential alternative to the synthetic hormones millions of women take after reaching middle age. A paper describing the findings was published in Nature Communications.


Women going through menopause, as well as those who have undergone cancer treatment or had their ovaries removed for medical purposes, lose the ability to produce important hormones, including estrogen and progesterone. Lower levels of these hormones can affect a number of different body functions. To counteract unpleasant symptoms, many women turn to combinations of hormone replacement medications—synthetic estrogen and progestin. Pharmacologic hormone replacement therapy (pHRT) with estrogen alone or estrogen and progestogens is known to effectively ameliorate the unpleasant symptoms. But hormone replacement carries an increased risk of heart disease and breast cancer, so it’s not recommended for long-term use. In these circumstances artificial ovaries could be safer and more effective.


Regenerative medicine approaches that use cell-based hormone replacement therapy (cHRT) offer a potential solution to temporal control of hormone delivery and the ability to restore the HPO (Hypothalamo-Pituitary-Ovarian) axis in a way not possible with pHRT. Scientists have previously described an approach to achieve microencapsulation of ovarian cells that results in bioengineered constructs that replicate key structure-function relationships of ovarian follicles as an approach to cHRT. In the present study the scientists have adapted an isogeneic cell-based construct to provide a proof-of-concept for the potential benefits of cHRT.


Tissue or cell encapsulation may offer effective strategies to fabricate ovarian constructs for the purpose of fertility and/or hormone replacement. Approaches using segmental ovarian tissue or whole-follicle implantation (typically with a focus on cryopreservation of the tissue for reproductive purposes) have resulted in detectable hormone levels in the blood after transplantation. Previous studies have also shown that autotransplantation of frozen-thawed ovarian tissue can lead to hormone secretion for over 5 years in humans.


Although these approaches can be used to achieve the dual purpose of fertility and hormone replacement in premenopausal women undergoing premature ovarian failure, they would have limited application in postmenopausal women who only need hormone replacement to manage menopausal symptoms and in whom fertility is not desirable. In full development, the technology described in this research is focused on hormone replacement, would meet the needs of the latter group of women that is the postmenopausal women.


The cell-based system of hormone replacement described in this report offers an attractive alternative to traditional pharmacological approaches and is consistent with current guidelines in the U.S. and Europe recommending the lowest possible doses of hormone for replacement therapy. In the present research sustained stable hormone release over the course of 90 days of study was demonstrated. The study also demonstrated the effective end-organ outcomes in body fat composition, uterine health, and bone health. However, additional studies will be required to determine the sustainability of the hormone secretion of the constructs by measuring hormone levels from implanted constructs for periods longer than 3 months in the rat model.


This study highlights the potential utility of cHRT for the treatment and study of conditions associated with functional loss of the ovaries. Although longer-term studies would be of future interest, the 90-day duration of this rodent model study is consistent with others investigating osteoporosis in an ovariectomy model. However, this study provides a proof-of-concept for cHRT, it suffers the limitation that it is only an isogeneic-based construct implantation. Scientists think that further studies in either allogeneic or xenogeneic settings would be required with the construct design described in this report in the path towards clinical translation given that patients who would receive this type of treatment are unlikely to have sufficient autologous ovarian cells for transplantation.





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


A mutated gene called RAS gives rise to a signalling protein Ral which is involved in tumour growth in the bladder. Many researchers tried and failed to target and stop this wayward gene. Signalling proteins such as Ral usually shift between active and inactive states.


So, researchers next tried to stop Ral to get into active state. In inacvtive state Ral exposes a pocket which gets closed when active. After five years, the researchers found a small molecule dubbed BQU57 that can wedge itself into the pocket to prevent Ral from closing and becoming active. Now, BQU57 has been licensed for further development.


Researchers have a growing genetic data on bladder cancer, some of which threaten to overturn the supposed causes of bladder cancer. Genetics has also allowed bladder cancer to be reclassified from two categories into five distinct subtypes, each with different characteristics and weak spots. All these advances bode well for drug development and for improved diagnosis and prognosis.


Among the groups studying the genetics of bladder cancer are two large international teams: Uromol (named for urology and molecular biology), which is based at Aarhus University Hospital in Denmark, and The Cancer Genome Atlas (TCGA), based at institutions in Texas and Boston. Each team tackled a different type of cancer, based on the traditional classification of whether or not a tumour has grown into the muscle wall of the bladder. Uromol worked on the more common, earlier form, non-muscle-invasive bladder cancer, whereas TCGA is looking at muscle-invasive bladder cancer, which has a lower survival rate.


The Uromol team sought to identify people whose non-invasive tumours might return after treatment, becoming invasive or even metastatic. Bladder cancer has a high risk of recurrence, so people whose non-invasive cancer has been treated need to be monitored for many years, undergoing cystoscopy every few months. They looked for predictive genetic footprints in the transcriptome of the cancer, which contains all of a cell’s RNA and can tell researchers which genes are turned on or off.


They found three subgroups with distinct basal and luminal features, as proposed by other groups, each with different clinical outcomes in early-stage bladder cancer. These features sort bladder cancer into genetic categories that can help predict whether the cancer will return. The researchers also identified mutations that are linked to tumour progression. Mutations in the so-called APOBEC genes, which code for enzymes that modify RNA or DNA molecules. This effect could lead to cancer and cause it to be aggressive.


The second major research group, TCGA, led by the National Cancer Institute and the National Human Genome Research Institute, that involves thousands of researchers across USA. The project has already mapped genomic changes in 33 cancer types, including breast, skin and lung cancers. The TCGA researchers, who study muscle-invasive bladder cancer, have looked at tumours that were already identified as fast-growing and invasive.


The work by Uromol, TCGA and other labs has provided a clearer view of the genetic landscape of early- and late-stage bladder cancer. There are five subtypes for the muscle-invasive form: luminal, luminal–papillary, luminal–infiltrated, basal–squamous, and neuronal, each of which is genetically distinct and might require different therapeutic approaches.


Bladder cancer has the third-highest mutation rate of any cancer, behind only lung cancer and melanoma. The TCGA team has confirmed Uromol research showing that most bladder-cancer mutations occur in the APOBEC genes. It is not yet clear why APOBEC mutations are so common in bladder cancer, but studies of the mutations have yielded one startling implication. The APOBEC enzyme causes mutations early during the development of bladder cancer, and independent of cigarette smoke or other known exposures.


The TCGA researchers found a subset of bladder-cancer patients, those with the greatest number of APOBEC mutations, had an extremely high five-year survival rate of about 75%. Other patients with fewer APOBEC mutations fared less well which is pretty surprising.


This detailed knowledge of bladder-cancer genetics may help to pinpoint the specific vulnerabilities of cancer cells in different people. Over the past decade, Broad Institute researchers have identified more than 760 genes that cancer needs to grow and survive. Their genetic map might take another ten years to finish, but it will list every genetic vulnerability that can be exploited. The goal of cancer precision medicine is to take the patient’s tumour and decode the genetics, so the clinician can make a decision based on that information.




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Emerging STAR in Molecular Biology, Synthetic Virology and Genomics: Clodagh C. O’Shea: ChromEMT – Visualizing 3D chromatin structure


Curator: Aviva Lev-Ari, PhD, RN


On 8/28/2017, I attend and covered in REAL TIME the CHI’s 5th Immune Oncology Summit – Oncolytic Virus Immunotherapy, August 28-29, 2017 Sheraton Boston Hotel | Boston, MA


I covered in REAL TIME this event and Clodagh C. O’Shea talk at the conference.

On that evening, I e-mailed my team that

“I believe that Clodagh C. O’Shea will get the Nobel Prizebefore CRISPR


11:00 Synthetic Virology: Modular Assembly of Designer Viruses for Cancer Therapy


Clodagh O’Shea, Ph.D., Howard Hughes Medical Institute Faculty Scholar; Associate Professor, William Scandling Developmental Chair, Molecular and Cell Biology Laboratory, Salk Institute for Biological Studies

Design is the ultimate test of understanding. For oncolytic therapies to achieve their potential, we need a deep mechanistic understanding of virus and tumor biology together with the ability to confer new properties.

To achieve this, we have developed

  • combinatorial modular genome assembly (ADsembly) platforms,
  • orthogonal capsid functionalization technologies (RapAd) and
  • replication assays that have enabled the rational design, directed evolution, systematic assembly and screening of powerful new vectors and oncolytic viruses.


Clodagh O’Shea’s Talk In Real Time:

  • Future Cancer therapies to be sophisticated as Cancer is
  • Targer suppresor pathways (Rb/p53)
  • OV are safe their efficacy ishas been limited
  • MOA: Specify Oncolytic Viral Replication in Tumor cells Attenuate – lack of potency
  • SOLUTIONS: Assembly: Assmble personalized V Tx fro libraries of functional parts
  • Adenovirus – natural & clinical advantages
  • Strategy: Technology for Assmbling Novel Adenovirus Genomes using Modular Genomic Parts
  • E1 module: Inactives Rb & p53
  • core module:
  • E3 Module Immune Evasion Tissue targeting
  • E4 Module Activates E2F (transcription factor TDP1/2), PI3K
  • Adenovirus promoters for Cellular viral replication — Tumor Selective Replication: Novel Viruses Selective Replicate in RB/p16
  • Engineering Viruses to overcome tumor heterogeneity
  • Target multiple & Specific Tumor Cel Receptors – RapAd Technology allows Re-targeting anti Rapamycin – induced targeting of adenovirus
  • Virus Genome: FKBP-fusion FRB-Fiber
  • Engineer Adenovirus Caspids that prevent Liver uptake and Sequestration – Natural Ad5 Therapies 
  • Solution: AdSyn335 Lead candidat AdSyn335 Viruses targeting multiple cells
  • Engineering Mutations that enhanced potency
  • Novel Vector: Homes and targets
  • Genetically engineered PDX1 – for Pancreatic Cancer Stroma: Early and Late Stage
On Twitter:

Engineer Adenovirus Caspids prevent Liver uptake and Sequestration – Natural Ad5 Therapies C. O’Shea, HHDI

Scientist’s Profile: Clodagh C. O’Shea


BS, Biochemistry and Microbiology, University College Cork, Ireland
PhD, Imperial College London/Imperial Cancer Research Fund, U.K.
Postdoctoral Fellow, UCSF Comprehensive Cancer Center, San Francisco, U.S.A


O’Shea Lab @Salk


  • 2016 Howard Hughes Medical Institute Faculty Scholar
  • 2014 W. M. Keck Medical Research Program Award
  • 2014 Rose Hills Fellow
  • 2011Science/NSF International Science & Visualization Challenge, People’s Choice
  • 2011 Anna Fuller Award for Cancer Research
  • 2010, 2011, 2012 Kavli Frontiers Fellow, National Academy of Sciences
  • 2009 Sontag Distinguished Scientist Award
  • 2009 American Cancer Society Research Scholar Award
  • 2008 ACGT Young Investigator Award for Cancer Gene Therapy
  • 2008 Arnold and Mabel Beckman Young Investigator Award
  • 2008 William Scandling Assistant Professor, Developmental Chair
  • 2007 Emerald Foundation Schola


Clodagh C. O’Shea: ChromEMT: Visualizing 3D chromatin structure and compaction in interphase and mitotic cells | Science


Clodagh C. O’Shea

In Press

Jul 27, 2017 – Salk scientists solve longstanding biological mystery of DNA organization

Sep 22, 2016 – Clodagh O’Shea named HHMI Faculty Scholar for groundbreaking work in designing synthetic viruses to destroy cancer

Oct 05, 2015 – Clodagh O’Shea awarded $3 million to unlock the “black box” of the nucleus

Aug 27, 2015 – The DNA damage response goes viral: a way in for new cancer treatments

Apr 12, 2013 – Salk Institute promotes three top scientists

Oct 16, 2012 – Cold viruses point the way to new cancer therapies

Aug 25, 2010 – Use the common cold virus to target and disrupt cancer cells?

Oct 22, 2009 – Salk scientist receives The Sontag Foundation’s Distinguished Scientist Award

May 15, 2008 – Salk scientist wins 2008 Beckman Young Investigator Award

Mar 24, 2008 – Salk scientist wins 2007 Young Investigator’s Award in Gene Therapy for Cancer

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Ido Sagi – PhD Student @HUJI, 2017 Kaye Innovation Award winner for leading research that yielded the first successful isolation and maintenance of haploid embryonic stem cells in humans.

Reporter: Aviva Lev-Ari, PhD, RN


Ido Sagi – PhD Student, Silberman Institute of Life Sciences, HUJI, Israel

  • Ido Sagi’s research focuses on studying genetic and epigenetic phenomena in human pluripotent stem cells, and his work has been published in leading scientific journals, including NatureNature Genetics and Cell Stem Cell.
  • Ido Sagi received BSc summa cum laude in Life Sciences from the Hebrew University, and currently pursues a PhD at the laboratory of Prof. Nissim Benvenisty at the university’s Department of Genetics in the Alexander Silberman Institute of Life Sciences.

The Kaye Innovation Awards at the Hebrew University of Jerusalem have been awarded annually since 1994. Isaac Kaye of England, a prominent industrialist in the pharmaceutical industry, established the awards to encourage faculty, staff and students of the Hebrew University to develop innovative methods and inventions with good commercial potential, which will benefit the university and society.

Publications – Ido Sagi

Comparable frequencies of coding mutations and loss of imprinting in human pluripotent cells derived by nuclear transfer and defined factors.
Cell Stem Cell 2014 Nov 6;15(5):634-42. Epub 2014 Nov 6.
The New York Stem Cell Foundation Research Institute, New York, NY 10032, USA; Naomi Berrie Diabetes Center & Department of Pediatrics, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA. Electronic address:

November 2014


Stem cells: Aspiring to naivety.
Nature 2016 12 30;540(7632):211-212. Epub 2016 Nov 30.
The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
November 2016

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Other related articles on Genetic and Epigenetic phenomena in human pluripotent stem cells published by LPBI Group can be found in the following e-Books on

e-Books in Medicine

9 results for Kindle Store : “Aviva Lev-Ari”

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    Etiologies of Cardiovascular Diseases: Epigenetics, Genetics and Genomics

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    Cancer Therapies: Metabolic, Genomics, Interventional, Immunotherapy and Nanotechnology in Therapy Delivery (Series C Book 2)

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    Regenerative and Translational Medicine: The Therapeutic Promise for Cardiovascular Diseases

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    Cardiovascular Original Research: Cases in Methodology Design for Content Co-Curation: The Art of Scientific & Medical Curation

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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.



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LIVE – Afternoon Session added – Gene Therapy Development & & Cell Therapy Bioprocessing – Overcoming Scientific and Development Challenges @Biotech Week Boston, October 5, 2016 Boston Convention and Exhibition Center





Leaders in Pharmaceutical Business Intelligence (LPBI) Group, Boston


will cover in REAL TIME

Biotech Week Boston, October 5, 2016 @ Boston Convention and Exhibition Center


In Attendance, streaming LIVE using Social Media

Aviva Lev-Ari, PhD, RN



Gene Therapy Development & Production


Gene Therapy Development & Production

8:25 am 5 mins

Chairperson’s Opening Remarks

8:30 am 30 mins

Extended Range and Next Generation Chimeric Antigen Receptor T Cells

  • Bruce Levine, Ph.D., Barbara and Edward Netter Associate Professor in Cancer Gene Therapy, University of Pennsylvania


  1. Lymphoeid Leukemia –  Persisting CTL019 Cells remains – Pediatrics ALL – Relapsed and Refractory B-celll
  2. Bedside back to Bench lessons: Days post infusion
  3. 28 days vs 51 cells
  4. quality of cells: related to Clinical Response
  5. 93% CR rate for r/r ALL after CTL019
  6. Novartis and UPenn Cell Center Building


  1. Diffuse Large B Cell Lymphoma
  2. Engineered T Cells and CHeckpoint ANtibody Therapies: Potential Synergies
  3. Days post-tumor injection: Non-responding patients: infusion of PD1 Inhibitor
  4. After treatment: Durable responses necrosis no tumor left
  5. Single Arm, Open-Label


  1. CART19
  2. CART BCMA – cells for Multiple Myeloma – Bone Marrow: Pre-treatment 70% myeloma cells
  3. Ibrutinib enhances chimeric antigen receptor T-cell engraftment and efficacy in Leukemia
  4. Synthetic Biology – Penn Platform Technology: Academic CAR Clinical Trials
  • Academic-Industry COllaboration with Novartis: Human Cell Therapy: 1996-2011
  • FromGene therapy to >>> Immunotherapy, to >>>>>> Gene Editing
  • TMUNITY – new mission with Leukmia and Lymphoma Foundation

9 am30 mins

Novartis Pharma: Stabilizing Lentiviral (LV) vector formulation for CART Application

  1. LV development of Vector Production – for Ex-vivo Therapy
  2. mamamalian cells + plasmid
  3. high titer, sequences liability &FTO
  • aggregation
  • Trunsduction
  • purification –
  • Phys-chem profile,
  • stable – – Stability in buffers for recovery of infectious particles – Purification
  • low cost
  1. LV: Optimize transient production:
  • FORMULATION SCEEEN – Preformulation of LVs: Drug DIscovery
  1. High throughput screen
  2. stability promoting buffers
  3. stability in process intermediates
  4. Attributes: Preformulation – success criteria – stable in transduction media/conditions’minimal loss of infectious particles
  5. LV with Transgene 1Preformulation: Effect of buffer and salts – 96 different conditions (buffer +salts)
  6. LV Aggregation & Loss of Infectivity – Accelerated Stree Studies
  7. Size Interpretations – Dynamic Light scattering (DLS): Buffer A, E, H
  8. Robustness: Preformulation buffers: Small, Medium, Large Rh:
  • pH 6.0 – 8.5 – X axis and
  • Salt 0 – 200nM – V Axis

9. Freeze-Thaw Studies – Accelerated Stress Studies: w/o Carbohydrate

  •  number of Freeze-thaw cycles X axis vs
  • % Control to standard – V Axis  

10, Infectious Titer: Primary T Cells

Case study: LV with Transgene #2 –

  • Platform formulation for LV is possible
  • Transduction in primary T cells showed high titer and minimal toxicity
  • stability studies of in-process intermediates and LV-DS provided additional evidence
  • 4 promising conditions identified for preformulation of LV vectors from HTS screens

 9:30 am 30 mins

Mesenchymal Stem Cells (MSCs) on Steroids

  • Oren Levy, HMS, Brigham and Women’s Hospital
I. Mesenchymal Stem Cells – readily accessible potent immunomodulatory secretome already used in >600 clinical trials
  1. Uncontrolled cargo’Inefficient targeting to disease sites – A microparticle-based cellular
  2. Particle-in-a cell approach in Prostate Cancer: PSA – Thapsigargin – PLGA – Poly Lactic co Glycol Acid – encapsulated agent’The cellular carriers MSCs
  3. Glioblastoma, Prostate
  4. Drug-loaded MSCs kill prostate cancer cells
  5. Drug release from G114 MP-loaded cells
  6. MDA-MB; LNCaP – Coculture with pretreated MSCs
  7. Drug-loaded MSCs inhibit tumor growth: days post-inoculation vs Probabilirt of Tumor-free SUrvival vs. Tumor Volume
  8. Particle Payloading: Boosting Potency of Administrated Cell Populations – for Drug delivery
  9. Controlled inhibition of the MSC proinflammatory
  10. Donor variability leads to Functional Variability – variability between 7 donors
  11. Budesonide microparticles boosted MSC
  12. Improved imaging of transplanted MSCs
  13. Retention of PLGA
  • MSC: Therapy
  • Betacells – Islet transplantation Macrophages – immunotherpy

II.  Targeting cells to disease site – Surface of MSC – lacks standard “homing”

  • tetharing – rolling – firm adhesion – transmigration- endothelial cells, basement membrane: Healthy vs Inflammed
  • lucocytes
  • Controlling MSC fate via small molecule pretreatment
  • RNA transfection
  • Bone marrow: PSGL-1/SLex MSCs exhibit enhanced homing to healthy and irradiated bone marrow
  1. A multi-step screening platform for identification of small molecules that improve MSC homing: Surface expression, FIrm adhesion, MSC homing
  2. A medium throughput screen identified Ro-31-8425
  3. Upregulated of CD11a, in response to pretreatment  increases MSC firm adhesion
  4. Pretreated MSCs exhibit superior therapeutic impact post systemic adm – suppression of local skin inflammation
  5. Ro-31-8425 Treated MScs va Vehicle treated MScs
  6. boosted clinical impacet in a MS Model
  7. Bradly applicable cell-based therapeutic platform
  8. mRNA transfection
  9. drug microparticles


10:30 am30 mins

Genetic Engineering Red Blood Cells for Therapeutic Function

    Robert J. Deans, Ph.D.,Chief Scientific Officer, Rubius Therapeutics
  • Rubius Therapeutics – Whitehead Institute/DARPA legacy
  • Flagship Venture founded
  • RCTs – development of ALternative Therapeutics
  • Platfoms: Transfusion /donor selection, risk of oncogenicity – uncontrolled division and secretion
  • Allogenic RCTs grown in Culture from Hemopoeitic Stem Cells (HSCs) In vitro and engineered to Purpose along the way
  • Expansion: CD34 + HSCs: Mobilized, cord blood
  • Cytoplasm: Native cytoplasmic, Tethered cytoplasmic , tethered surface native surface
  • Visualizing Red Cell Therapeutics – RCT during differentiation
  • Reliablly use cell surface markers to track the cells as hey differentiate
  • Flow Cytometry
  • Automated platform foe creating Master cell BioBank
  1. Apheresis Transduction
  2. clinical scale
  3. Rapid prototyping capability for candidate Selection, Vector generation, Human CD34+
  4. Prototyping for Pipeline
  5. Exogenous protein is retained: Cultured RBCs circulating in vivo like Human RBCs
  6. Experimental Setup: RCT circulation in similar to HuRBCs
  7. RCTs can possess antibody drugs on their surface for clearance and therapeutic action
  8. Target: scFv antiVirusAg – capture circulating antigen in vitro and in vivo – Virus Antigen – label antibody
  9. Phenylketonuria (PKU) – CNS morbidity from Serum PHE – it is easily measured, gold standard diagnostic and clinical biomarker
  • Novel protein, cell & gene therapy treatment
  • Existing PAL-RBCs have clinical grade potency at currently achievable manufacturing scale: Serum PHE vs Units PAL-RBS IRON
  • dose dependent PHE expression


11 am30 mins

Novel Approaches to Gene Editing and Gene Delivery 

In Vivo gene therapy

    Andrew Scharenberg, M.D.,Professor of Pediatrics, Adjunct Immunology / Co-director, Program in Cell and Gene Therapy, University of Washington / Seattle Children’s Research Institute
  1.  Viral vector tranfer
  2. cell mediated immunity
  3. Humoral immunity
  4. Evasion/tolerance – neonatal adm of vector
  5. Challenges: Unmet need Goal – Primary cell: DNA delivery to nucleus
  • Cells have mupliplr mechanisms to prevent foreign DNA from entering the nucleus
  • Viruses evade these mechanisms

Non-enveloped Virions – classic gene therapy vector AAV

  • Viral capsid serves multiple roles:
  • AAV has no mechanism to neutralize nuclear silencing mechanism

Enveloped Virions: Retro and Lenti-Virus

Re-dosable in vivo gene transfer

LNP’s mimic the endosomal uptake and escape functions of AAV Capsid

  • mRNA delivery to build a capsid “life boat” – reverse transcribe the gemone and send it in
  • RNA packing, minus strand systhesis, plus strand synthesis, budding, ER vesicular transport ENTRY repair

HBV-based Non-Viral RNA Gene Transfer System: Electric Poration (EP) or LNP

Pregenomic (pgRNA) Vector RNA architecture

  1. Viral RNA – Pol transfers to 3′ DR1 for _ strand sysnthesis
  2. Vector RNA – payload cap… pA
  3. pgRNA with GFP cassette flanked by SB Tn sites – Inhibition and Translation areas
  4. BFP vs GFP
  5. Validation of pgRNA reverse transcription: HepG2 cells transfected with indicated RNAs plus POL/CORE/X mRNAs, then cultured for 2 weeks
  6. Plasma DNA vs Tn-pgRNA, Tn-pgRNA + 5B vs COntrol
  7. pgRNA/SB achieves stable luciferase expression
  8. No-Viral RNA Gene Transfer System: Status and Future
  9. Encouraging evidence of functional synthetic recycling RT vector: SERT – Synthetic encoded reverse transcription)
  10. Transposon IR/DR – flaked pgRNA payload


11:30 am30 mins

Development of Stem Cell Derived Extracellular Vesicles into a Non-Living Regenerative Therapeutic Drug Candidate

    Keil  Ph.D., Capricor, Inc.
  1. Cardioshere-Derived Cells – DOnors of cells – Primary cardiac tissue’Key functions – paracrine:
  2. CDC Manufacturing
  3. DOnor Heart from organ procurement —
  4. Cell Therapy: Autologous vs Allogenics
  5. Cell engraftment
  6. Autologous vs Allogenics Clinical Trials:
  7. Caduceus vs Dynamic
  8. Scar tissue

What are exosomes? as a therapeutics

  • rich in RNA
  • CDC EVs: By the numbers
  • Exosomal Markers vs CDC Marker – CD 105

Acute MI Model

  • Viable mass increased, Myocyte size increased, EF increased,
  • Identified 146a as a major player in cardio-protection
  • CAP2003 manufacturing Process
  • CDC-EV Manufsacturing Overview vs CDC Product
  • Conditioned Medium collection
  • day 1 vs 15 days
  • Drug Product Formulation
  • Choosing a Formulation Buffer: pH
  • 30 days stability
  • Process Robustness: Particle Size and pH: 4 diffrent donors (variability is related to culture and to patients) – 10 formulation runs: EV size vs diafiltrate pH (average =7)
  • Process Robustness: Concentration and Cargo
  • Human mir panel v3.0
  • nCounter miRNA analysis
  • Particle foes not increase linearly with protein
  • Process Robustness: miRNA 146 & 210
  • Process Impurities & Residual: Residual  Fibronectin in CM vs BSA wash-out
  • CDC-EVs have anti-apoptotic, anti fibrotic, immunomodulatory and pro-regenerative activities similar to CDCs
  • New indications
  • Pathway to iND: Pre-Clinical Efficacy
  • Average Total clinical Score
  1. conjunctival injection
  2. corneal opacification
  3. corneal area affected
  4. corneal neovascularization
  5. aqueous flare
  6. Ongoing Pre-Clinical: biodistribution,
  7. Upscaling Exosomes: Bioreactor CDCs
  8. Current Process vs Target Process

Next Gen Storage: Lyophilization (Astraunatu EV

State of Capricor’s Art


Cell Therapy Bioprocessing – Innovations in Cell Processing – Part 2


2:10 pm 5 mins

Chairperson’s Remarks


2:15 pm30 mins

CAR T production in G-Rex: Less is More

    Juan Vera, M.D.Associate professor Center for Cell and Gene Therapy, Baylor College of Medicine., Baylor College of Medicine
  • Adoptive T-cells Transfer
  • Donor Lymphocytes blood draw — antigen specificities == cell expansion == infusion back to aptient – antigen specific T-cells to iincrease immunogenecity
  • Low seeding density results in greater fold expansion
  • Low cell density + Greater cell expansion: Optimal time for cell harvest vs maximum cell density
  • Volume of Media vs Cells density
  • Upfront media addition resulted in shortest culture period
  • Conventional cultureware  – risk for contamination
  • G-Rex – Plate vs G-Rex – G-REX 5; G-Rex 100; G-REX -500 (flacks)
  • superior cellnumber not due to increase cell number – improved cell output
  1. phase 1 – what is optimal seeding density – cells expansion in 12 days
  2. phase 2 – optimal volume in media – I=1L
  3. phase 3 –
  • are observations reproducible?
  • Cell harvest with The GatheRex – in 5 minutes you collect one billion cells (density
  • CAR-T cells: Preparation of genetically-modified T cells
  • PSCA – solid Tumor
  • NT T cells – suppress tumor cells by immunoresponse to antigen specific to CAR-T
  • CAR-T cell expansion in G-REX – day in culture vs Glucose concentration – inverse correlation
  • Cell performance of production of CAR=t in G-REX
  • Phynotype of CAR-T cells: 24 wells vs G-REX
  • CCR7 vs CD62L: Conventional vs G-REX  – superior Phynotype of CAR-T Cells expanded inside G-REX
  • stimulation beads = not used i=with G-REX
  • media vs cell ratio – serology
  • Persistance of CAR-T cells 6 month post transfer – rechallenge the cells ramping up withstand multiple re-challenge
  • Post transfection – transduction 3 days after

2:45 pm40 mins

Approaches for Determining Technologies to Pursue for Cell Therapy Bioprocessing

  • Brian Murphy, Ph.D., Director of Development, Celgene Cellular Therapeutics

Celgene cellular therapy

  • PDA-002 -MSC
  • PNK-007 – NK cells
  • TST-001
  • GM-
Technologies – CMC Feasibility: make enough cells, the right phynotypes make them GMP – secure cell supply
  • preclinical
  • Phase 1
  • phase 2
  • phase 3 – commercial: Controllable, scaleable, compliant, cost-effective

Scale up: Phase appropriate under scale or over scale vs utilization

  • TIPS: Cell experience need be controlled
  • Static culture technology vs suspension culture
  • Stability in media
  • metabolite
  • Biology Informs Technology: Control critical parameters vs Failure mode
  • After Biology – other factors:
  1. cost
  2. capacity

PDA-002 – Two tier banking system for Cell therapies – adherent cells grown in suspension or microcarriers

Growth on microcarriers in Bioreactor

  • innoculation efficacy
  • harvest formulation
  • fill
  • cryopresevation drug substance

Phase 2 – Process technology

Phace 3 – Process technology for Dose increase — Drug Product — Shipping Product

Biological needs of the cels: Day 11 Media x vs Media Y

Impact of a 35 day Culture — long process: Process duration, Trouble shooting Characterization:

  • 7 days,
  • 21 days
  • 35 days


PNK-007: Process Overview – sensitive at few time points in while grown in suspension

Supply Projection & Inventory: 1,2,3 years

Phase 1

phase 2

Phase 3

Conclusion Multiple Groups involved in Production

  • Bioprocess development
  • Stem cell research
  • Analytical development
  • Manufacturing Operations
  • Quality Assurance

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Cell & Gene Therapy BioProcessing & Commercialization

Cell Therapy Bioprocessing – Innovations in Process Technologies, Friday, October 7, 2016, Boston Convention and Exhibition Center

Informa Life Sciences lead the market in providing quality, expert-led conferences; delivering the expert knowledge our clients need to excel in their professional roles and guaranteeing a competitive advantage for their organisations.

Our diverse portfolio covers the Pharmaceutical (Drug Discovery, Clinical Development, Regulatory Affairs, Biopharmaceutical, Generics and Business Strategy), Medical Devices and Diagnostics, Fine Chemicals and Agrochemicals and Veterinary Medicine arenas

8 am 5 mins

Chairperson’s Remarks

  • Jon A. Rowley, Ph.D., RoosterBio Inc.

10 am 30 mins

Applications for Use of the Lovo Cell Processing System in Cell Therapy Process Development and Manufacturing

  • Alaina C. Schlinker, Ph.D., Fresenius Kabi USA, LLC

10:30 am 30 mins


  • Philip G. Vanek, Ph.D., GE Healthcare

11 am 30 mins

Platform Processes for PSC Derived Product Manufacture

  • Nick Timmins

11:30 am 30 mins

Building the 3rd Pillar of Medicine: Bioprocessing for Cell Therapies

  • Steve K.W. Oh, Bioprocessing Technology Institute

12 pm 45 mins

Enabling Technologies for Efficient Downstream Processing of AAV Viral Vectors

  • Orjana Terova, Purification, Thermo Fisher Scientific

1:55 pm 5 mins

Chairperson’s Remarks

  • Dominic Clarke, Charter Medical

2 pm 30 mins

Innovative Device Development for Cell Therapies

  • Jamie Piret, Sc.D., The University of British Columbia

2:30 pm 30 mins

Applying Single-Use Technologies

  • Paula Alves, Ph.D., iBET, Portugal

3 pm 30 mins

Manufacturing Multiple Cell Based Products Simulaneously

  • Gail K. Naughton, Ph.D., Histogen Inc.

3:30 pm 30 mins

Scale Out iPSC Derivation and Differentiation Processes for Cell Therapy

  • Wen Bo Wang, Ph.D., Cellular Dynamics International, a FujiFilm company


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