Simple Complexity in an Evolving World: Rising to the Challenge

noisymaniacalBiotechnology

Feb 20, 2013 (4 years and 1 month ago)

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DCGT research priorities:

Led by Critical Path Challenges

Virology

Retroviruses, lentivirus, adeno, filovirus

Immunology

Anti
-
viral immunity, immunobiology of cell therapy and
xenotransplantation

Cell biology

Control of differentiation in animal models,

stem cell biology

Cancer biology

Antigen characterization, Molecular biomarkers,

animal models

Biotechnology

-

Genomics, flow cytometry, proteomics

Tissue Safety Program

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Division of Cellular and Gene Therapies

Raj Puri, Ph.D., M.D., Division Director

Kimberly Benton, Ph.D., Deputy Director


Division of Cellular and Gene Therapies

Raj Puri, Ph.D., M.D., Division Director

Kimberly Benton, Ph.D.,

Deputy Director


Cellular and Tissue

Therapy Branch

Steven Bauer,
Ph.D
., Chief



Tumor Vaccines and

Biotechnology Branch

Raj Puri, Ph.D., M.D., Chief



Cell Therapies Branch

Keith Wonnacott, Ph.D., Chief



Gene Therapies Branch

Daniel Takefman, Ph.D., Chief



Gene Transfer and

Immunogenicity Branch

Andrew Byrnes, Ph.D., Chief


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Tumor Vaccines and

Biotechnology Branch (TVBB)

Raj K. Puri, M.D., Ph.D.

Cancer Biology and Genomics Program


Syed R. Husain, Ph.D., Staff Scientist

Bharat H. Joshi, Ph.D., Staff Scientist


Michail Alterman, Ph.D.


Proteomics Program


Shyh
-
Ching Lo, M.D., Ph.D.

Tissue safety Program



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Scientific and Regulatory
Challenges in cancer Vaccines and
Immunotherapy


Puri lab


Complex biology of cancer


Identification of appropriate targets and
specificity of target


Appropriate tests and biomarkers for identity and
potency of cancer therapeutics/vaccines


Animal models


safety and efficacy


Immune biomarkers of response and disease
monitoring


5

Research programs addressing

scientific and regulatory challenges


Specific Programs:



Characterization of tumor associated cell surface
proteins (antigens or receptors) for identity and potency
and target these receptors for cancer therapy



Animal models of human cancer to assess safety and
efficacy of tumor targeted agents including cancer
vaccines, immunotoxins, cell and gene therapy products



Characterization of cancer vaccines and embryonic stem
cells by genomics technology to identify cancer stem
cells and biomarkers of identity and potency and
response to cancer vaccines


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Characterization of tumor associated cell
surface receptors and antigens


Discovery of overexpression of Th
-
2 derived cytokine
receptors in tumors



Interleukin
-
4 receptors


RCC, malignant glioma, AIDS
-
Kaposi’s sarcoma, colon
cancer, breast carcinoma, NSCLC, prostate cancer, ovarian
carcinoma, mesothelioma, hematological malignancies
(CLL
-
B),

PDA, biliary ductal cancer and SCCHN


Interleukin
-
13 receptors


RCC, malignant glioma, pediatric glioma, AIDS
-
Kaposi’s
sarcoma, SCCHN, ovarian carcinoma, PDA,
pheochromocytoma, brain stem glioma and prostate cancer


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Animal models of human cancer to assess

safety, toxicity and efficacy of cancer

targeted agents


Ovarian Cancer


Pancreatic cancer


Glioblastoma multiforme


SCCHN


Mesothelioma


Lung cancer





AIDS
-
Kaposi’s tumors


Breast cancer


Hodgkin’s lymphoma


Melanoma


Pheochromocytoma

Pheochromocytoma

8

Conclusions and Ongoing Projects


Certain human tumors express high levels of IL
-
4R and IL
-
13R
in vitro

and
in vivo;
the structure and signal transduction
through these receptors is different in cancer and normal
cells.



IL
-
4R and IL
-
13R can be effectively targeted by immunotoxins
by themselves or when combined with other agents for cancer
therapy.



IL
-
13R

2 is involved in PDA invasion and metastasis and
may be a prognostic biomarker of disease.



IL
-
13R

2 can signal through AP
-
1 pathway and regulated by
Adrenomedullin
in vitro

and
in vivo.



IL
-
13R

2 is a novel tumor rejection antigen
-

IL
-
13R

2 cDNA
vaccination induces immune response in preventive and
therapeutic murine cancer models.

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Analytical proteomics for the characterization
of biologic products and discovery of
biomarkers

Michail Alterman, Ph.D.

Senior Investigator,

Tumor Vaccines and Biotechnology Branch,

Division of Cellular and Gene Therapies


Laboratory Goal:



Development and application of mass spectrometry
-
based proteomic tools for qualitative and
quantitative measurement of cellular products, cell
substrates and vaccines and discovery of potential
biomarkers of cancer






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Mission relevant questions to be impacted by
application of proteomics tools


Implementation of new better, clearer and more defined means
for regulatory decision making in evaluation and review of cell
substrates, vaccine products and other biologics


Development of valid protein biomarkers for examining drug
safety and efficacy through prediction of therapeutic effect,
selection of appropriate dose, and identification of risk of
toxicity.


Development of improved strategies for biotechnology product
physicochemical comparability assessments


Development of effective tools to be used in biologics and
protein therapeutics manufacturing process controls and
release testing


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Specific aims


and achievements since Site Visit in April 2010

Specific Aim # 1.

Development of mass spectrometry
-
based analytical
tools for testing of biological product quality and identity.


Manuscript published in Analytical Biochemistry


Specific Aim # 2.

Identification of proteomics
-
based cellular molecular
signatures that may be tested as predictors of therapeutic success
and be developed as biomarkers of safety and effectiveness in
preclinical animal models.


Invited talk at an international meeting, one poster presented at an
international meeting and one at a national meeting, two manuscripts
in preparation


Specific Aim #3.

Proteomics
-
based analysis of influenza virus and
vaccines.


One invited seminar, manuscript in preparation




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Conclusions


A new label free quantitation technique was developed
that allows determination of a ratio of co
-
migrating in gel
electrophoresis proteins


Two prospective protein groups were identified that
could be used as biomarkers for characterization of cell
substrates


MS
-
based proteomics methods can be used to
identify/distinguish different influenza variants and clades
and quantify proteins in flu vaccines and virus
preparations


A comprehensive proteome map of human MSC is under
development; a number of prospective biomarkers of cell
sub
-
culturing was found among >4000 identified proteins


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Ongoing projects


1.
Development of a new potency/stability
assay for flu vaccine.


2.
MSC project:





i. identify protein signatures characteristic
for MSC from multiple donors


ii. identify molecular signatures that could
be used to discriminate product that
produced a desirable clinical outcome


Tissue Microbiology Laboratory

for Safety of Human Tissues

Intended for Transplantation




Shyh
-
Ching Lo, Ph.D., M.D.








SV Report (2010) for The Tissue Laboratory Program
of Tumor Vaccines and Biotechnology Branch


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Public Health, Regulatory Challenges and

Critical Path Scientific Research:


To Better Understand The Scientific Trends and Further
Strengthen The Dialogue with The Stakeholders of Human
Tissue


Increasing number of tissues are recovered and processed as
grafts by the industry and used by the medical practice each
year.



The OCTGT within CBER regulates human cells, tissues, and
cellular and tissue
-
based products (HCT/Ps) in order to
prevent the spread or transmission of communicable
diseases.



The scientific capability of the new program needs to cover a
wide spectrum of expertise with very different microbial
pathogens, including bacteria, fungi, viruses and protozoa
parasites.



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Research Programs of Tissue
Laboratory




Specific Aims:


To establish the required scientific capabilities to directly
support regulatory needs for tissue safety



To adopt new technologies for rapid detection of infectious
pathogens with high sensitivity in human tissues being
processed for transplantation



To evaluate innovative methods for effective inactivation of
various pathogens while preserving tissue quality for clinical
needs



To explore new scientific approaches for detection and
characterization of previously unknown or newly emerging
infectious pathogens that would likely threaten the safety of
human tissue grafts





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Laboratory research projects initiated
to accomplish the specific aims:


Establishment of the laboratory capability of clinical microbiology
for detection and characterization of microbes with tissue safety
concerns

--

Investigation of incidents and validation of industrial
sterilization methods



Development of rapid, highly sensitive technology for detection of
pathogens in tissues intended for transplantation
--

R
eal
-
time
qPCR arrays of the targeted high
-
risk bacteria and RCDAD viruses



Development of high
-
throughput genomic sequencing capability
for detection and characterization of previously unknown
infectious pathogens in tissue or tissue
-
based products
--

Readiness of scientific capability against potential newly emerging
threats of public health



Identification of “biomarkers” associated with the injury
mechanisms in cell/tissue and development of the injury
-
associated microarray
--

Assessment of the degree of tissue injury
in pathogen inactivation using various chemical treatments

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Conclusions and Ongoing Projects


A new tissue microbiology laboratory has been
established under the DCGT and the DHT to
enhance both the safety and the availability of
high quality human tissue intended for
transplantation.



Development of New Technologies and
evaluation of innovative methods is being
implemented to guard the safety and quality of
tissue intended for transplantation





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Thank you