# Presentation: 126323

## Office of Biotechnology Products

**Elizabeth (Wendy) Shores**
** ****Deputy Director, OBP**

- July 26, 2007
- Advisory Committee Meeting

**RESEARCH**

**REVIEW &**
**REGULATION**

## Center for Drug Evaluation and ResearchOffice of Pharmaceutical  ScienceOBP

**Notes:**
This is an abbreviated organizational chart to illustrate the general structure of the Division and to show the relative positions of those being reviewed today within that structure.
Dr. Bonvini is serving as the Acting Deputy Director of the Division and Dr. Patrick Swann is in charge of the full-time reviewers in the Regulatory Science & Policy Branch.
The 3 research labs are here headed by Dr. Clouse, Dr. Bonvini, and Dr. Kozlowski.
Dr. Markovic is a Staff Fellow in Dr. Clouse’s lab and Dr. Frucht and Dr. Feldman are Staff Scientists in the LCB and LIB, respectively.

## OBP: What We Do

**	****Regulatory Review **
**	****	****Scrutinize Submissions from Industry**
** ****	****	****	****IND, IDE, 5(10)k**
**	****	****	****BLA, NDA (~90 approved), PMA**
**	****	****	****Supplements**
**	****	****Inspections (as product reviewer)**
** ****	****	****Guidance Documents**
** ****	****	****Meetings**
** ****	****	****Consults (area of expertise)**
**	****Bench Research**
**	****	****High Quality**
**	****	****Mission Relevant**

## Factors For Biotechnology Products

** ****Variety of Cell Substrates**
**	****Microbiology (virus, prion), Host Cell Molecules**
** ****Heterogeneity**
**	****Glycosylation, Oxidation, PEGylation**
** ****Higher Order Structure : Structure/Function**
**Receptor Interaction and Signaling**
** ****Manufacturability**
**	****Scale-Up, Solubility, Aggregation**
** ****Mechanism of Action: Often Unclear**
**	****Design of Bioactivity Assays**
** ****Immunogenicity: **
**	****Safety and Efficacy Issues**
** ****Comparability: Products Rarely Static**
** ****Counter Bioterrorism: Animal Rule, Stockpiles**

## Slide 5

## Slide 6

##  Immunity

** ****Immunity**

**Fc********RI,II,III**
**C1q**

- Jefferis et al.
-  *Immunology Letters*
- (2002) 82; 57-65.
- & PDB 2IG2, 1FC1

- Generation and Control of Antibody Responses to Proteins **[Ed Max and Ralph Bernstein]**

## Microbiology: Infection 

**Microbiology: Infection**

- Cytokines and Chemokines in HIV **[Michael Norcross and Jinhai Wang]**
- Characterization of Anthrax Lethal Toxin and Role in Disease **[David Frucht]**

## Innate Immunity &

**Innate Immunity &**
**  ****Soluble Mediators**

- Toll Like Receptor agonists
- 	and innate immunity **[Serge Beaucage]**
-     	* *

**Notes:**
The interpretation: 

	Any given GTP-binding protein, including Cdc42, has two nucleotide bound states.  These are GDP-inactive form and GTP active form.  The cycle between GDP and GTP is tightly regulated by the actions of GEF (Guanine nucleotide exchange factor), which promote GDP to GTP exchange and activate G-protein.  The second regulatory proteins are GAPs (GTPase activating protein), GAPs have the ability to accelerate intrinsic GTPase activity to hydrolyze GTP and bring Cdc42 back to inactive state.  The activity of Cdc42 can also be regulated by extracellular stimuli, for example, EGF-stimulated signaling.

	Once Cdc42 binds GTP, then it becomes active and talks to down-stream target proteins to regulate cellular functions, for example, cell growth regulation.  Like oncogene product Ras proteins, Cdc42 has the ability to induce cellular transformation when it is constitutively activated.

	Two different transformation assays are included in this slide, namely low serum growth and soft agar growth.  Both of them are in vitro transformation assays.  The first measures the lowered requirement for growth factors for transformed cells.  The second tests the less anchorage dependence for transformed cells.  NIH3T3 cell stably expressing either RasG12V or Cdc42F28L, both of them are constitutively activated, seeded under the experimental conditions.  The pictures were taken on the Day 6 for low serum growth or two weeks for soft agar growth.    

	The molecular mechanisms by which Cdc42–induced cellular transformation, we believe, is that the activated Cdc42 has the ability to inhibit Cbl-catalyzed ubiquitination and degradation of EGF receptor, resulting in the accumulation of EGF receptor in the cells and cellular transformation.

	Cdc42 has been found overexpressed in human breast cancers and implicated in cell invasiveness.  The EGF receptor is often overexpressed in breast cancer, but rarely amplified at the gene level.  In most cases, the mechanisms underlying the increased protein levels of receptor in human breast cancers are not known.  Now we try to understand whether the the activity status of Cdc42 is responsible for the elevated levels of EGF receptor in human breast cancers and use information we obtain from these studies to develop potential biomarkers and the treatment of human breast cancer. 

## Industrialization &

**Industrialization &**
**Assay **
**Development**

- Improved ODN Synthesis for Diagnostics and Therapeutics**            [Serge Beaucage]**

- Oligo Microarray

**Notes:**
The interpretation: 

	Any given GTP-binding protein, including Cdc42, has two nucleotide bound states.  These are GDP-inactive form and GTP active form.  The cycle between GDP and GTP is tightly regulated by the actions of GEF (Guanine nucleotide exchange factor), which promote GDP to GTP exchange and activate G-protein.  The second regulatory proteins are GAPs (GTPase activating protein), GAPs have the ability to accelerate intrinsic GTPase activity to hydrolyze GTP and bring Cdc42 back to inactive state.  The activity of Cdc42 can also be regulated by extracellular stimuli, for example, EGF-stimulated signaling.

	Once Cdc42 binds GTP, then it becomes active and talks to down-stream target proteins to regulate cellular functions, for example, cell growth regulation.  Like oncogene product Ras proteins, Cdc42 has the ability to induce cellular transformation when it is constitutively activated.

	Two different transformation assays are included in this slide, namely low serum growth and soft agar growth.  Both of them are in vitro transformation assays.  The first measures the lowered requirement for growth factors for transformed cells.  The second tests the less anchorage dependence for transformed cells.  NIH3T3 cell stably expressing either RasG12V or Cdc42F28L, both of them are constitutively activated, seeded under the experimental conditions.  The pictures were taken on the Day 6 for low serum growth or two weeks for soft agar growth.    

	The molecular mechanisms by which Cdc42–induced cellular transformation, we believe, is that the activated Cdc42 has the ability to inhibit Cbl-catalyzed ubiquitination and degradation of EGF receptor, resulting in the accumulation of EGF receptor in the cells and cellular transformation.

	Cdc42 has been found overexpressed in human breast cancers and implicated in cell invasiveness.  The EGF receptor is often overexpressed in breast cancer, but rarely amplified at the gene level.  In most cases, the mechanisms underlying the increased protein levels of receptor in human breast cancers are not known.  Now we try to understand whether the the activity status of Cdc42 is responsible for the elevated levels of EGF receptor in human breast cancers and use information we obtain from these studies to develop potential biomarkers and the treatment of human breast cancer.