Novel Technique for Early Detection of Pancreatic Cancer 7/28/10 Payam Chini, MD Rony Francois, MD/Ph.D. Candidate Francisco Delgado, Ph.D. Candidate Tal Salz, Ph.D. Candidate Process by which large quantities of antibodies (targeted against a particular antigen X) can be produced. A mouse is immunized by injection of an antigen X to stimulate the production of antibodies targeted against X. The antibody forming cells are isolated from the mouse's spleen. Monoclonal antibodies are produced by fusing single antibody-forming cells to tumor cells grown in culture. The resulting cell is called a hybridoma. Each hybridoma produces relatively large quantities of identical antibody molecules. By allowing the hybridoma to multiply in culture, it is possible to produce a population of cells, each of which produces identical antibody molecules. These antibodies are called "monoclonal antibodies" because they are produced by the identical offspring of a single, cloned antibody producing cell. Once a monoclonal antibody is made, it can be used as a specific probe to track down and purify the specific protein that induced its formation.

Pancreatic Cancer Mortality U.S. Cancer Statistics Working Group. United States Cancer Statistics: 1999–2006 Incidence and Mortality Web-based Report. Atlanta: U.S. Department of Health and Human Services, Centers for Disease Control and Prevention and National Cancer Institute; 2010. Available at: www.cdc.gov/uscs

Why pancreatic cancer has such a high mortality? Advanced stage of disease once diagnosed Most patients experience symptoms late in the course of disease

How do we diagnose pancreatic cancer? CT High sensitivity for tumors greater than 2.0cm Significant decline in sensitivity for smaller lesions Endoscopic Ultrasound More sensitive than CT for detecting smaller lesions Very operator dependent Difficult to identify lesions in setting of chronic pancreatitis MRI Less sensitive than CT and EUS for detecting Pancreatic Cancer

Screening for Pancreatic Cancer An ideal imaging protocol would identify pancreatic cancer early Allow implentation of a screening program for high risk population

Innovation Improve detection of Pancreatic Cancer Utilize tropism of oncolytic virus Nanotechnology Improve detection of Pancreatic Cancer

Normal Pancreatic Ductal Cells Pancreatic Cancer Cells Oncolytic Viruses Oncolytic Virus Newcastle Disease Virus (NDV) Selectively infects human cancer cells Anti-tumor activity in humans Normal Pancreatic Ductal Cells Pancreatic Cancer Cells

Nanoparticles Man-made structures on the scale of 1-100nm Unique properties not seen with materials on larger scales Favorable profile of interaction with cell surface

Formulated from highly cross-linkable polymers in variety of forms Nanoparticles Formulated from highly cross-linkable polymers in variety of forms Surface functionalization can create “targetable” nanoparticles Can be loaded with contrasts agents such as Gadolinium and Ferric ammonium citrate

Specific Aim 1 Develop MRI-detectable nanoparticles conjugated to antibodies against pancreatic tumor cell antigen for the purpose of early detection of pancreatic cancer MRI-detectable nanoparticles Antibody against malignant pancreatic cells

tumor cells that are associated with Newcastle Disease Virus Specific Aim 1.1- Identify and purify the surface antigens of pancreatic tumor cells that are associated with Newcastle Disease Virus Tumor antigen Mass spectrometry Immunoprecipitation http://www.springerprotocols.com/Full/doi/10.1385/159259655X:283?encCode=U09QOjM4MjpYLTU1Ni05NTI5NS0x&tokenString=HO9yQw0MDTE+r+qUV50lNQ==&access=denied

Immunization with Antigen Specific Aim 1.2: Generate pancreatic tumor specific antibodies and conjugate them to MRI-detectable nanoparticles Immunization with Antigen Myeloma cells plasma cells Monoclonal Ab Nano particle Process by which large quantities of antibodies (targeted against a particular antigen X) can be produced. A mouse is immunized by injection of an antigen X to stimulate the production of antibodies targeted against X. The antibody forming cells are isolated from the mouse's spleen. Monoclonal antibodies are produced by fusing single antibody-forming cells to tumor cells grown in culture. The resulting cell is called a hybridoma. Each hybridoma produces relatively large quantities of identical antibody molecules. By allowing the hybridoma to multiply in culture, it is possible to produce a population of cells, each of which produces identical antibody molecules. These antibodies are called "monoclonal antibodies" because they are produced by the identical offspring of a single, cloned antibody producing cell. Once a monoclonal antibody is made, it can be used as a specific probe to track down and purify the specific protein that induced its formation. http://www.accessexcellence.org/RC/VL/GG/monoclonal.php

Specific Aim 1.3: Test antibody-conjugated nanoparticles in vivo using pancreas cancer mouse model inject MRI Process by which large quantities of antibodies (targeted against a particular antigen X) can be produced. A mouse is immunized by injection of an antigen X to stimulate the production of antibodies targeted against X. The antibody forming cells are isolated from the mouse's spleen. Monoclonal antibodies are produced by fusing single antibody-forming cells to tumor cells grown in culture. The resulting cell is called a hybridoma. Each hybridoma produces relatively large quantities of identical antibody molecules. By allowing the hybridoma to multiply in culture, it is possible to produce a population of cells, each of which produces identical antibody molecules. These antibodies are called "monoclonal antibodies" because they are produced by the identical offspring of a single, cloned antibody producing cell. Once a monoclonal antibody is made, it can be used as a specific probe to track down and purify the specific protein that induced its formation. http://www.accessexcellence.org/RC/VL/GG/monoclonal.php

Conclusion and Future Studies Modalities for early detection of pancreatic cancer needed Our proposal Novel technique utilizing the properties of oncolytic viruses and nanoparticles Will lay the foundation for future studies that will test this technology in human subjects

Group Leaders: Acknowledgements Dr. Philipp Dahm, MD Dr. Keith Muller, Ph.D.

Questions? Process by which large quantities of antibodies (targeted against a particular antigen X) can be produced. A mouse is immunized by injection of an antigen X to stimulate the production of antibodies targeted against X. The antibody forming cells are isolated from the mouse's spleen. Monoclonal antibodies are produced by fusing single antibody-forming cells to tumor cells grown in culture. The resulting cell is called a hybridoma. Each hybridoma produces relatively large quantities of identical antibody molecules. By allowing the hybridoma to multiply in culture, it is possible to produce a population of cells, each of which produces identical antibody molecules. These antibodies are called "monoclonal antibodies" because they are produced by the identical offspring of a single, cloned antibody producing cell. Once a monoclonal antibody is made, it can be used as a specific probe to track down and purify the specific protein that induced its formation.