Chimeric Antigen Receptor

Gene Therapy For Cancer

Kevin HuginsDuy-Khiem Chanh Pham

The Beginning

Roblin and Friedmann, authors of “Gene therapy for human genetic disease?” in Science 1972

http://www.jstor.org/stable/1732705?seq=3

The first approved gene therapy in 1990 under Dr. Anderson on a 4-year old with ADA deficiency.

Advancement in Gene Therapy

The first efforts focused on modification of T-Cell receptors.

Problem: in vivo modified T-Cells were removed by negative selection by body’s immune system.

Solution: Create an entirely new receptor that could detect cancer cells but avoid negative selection. (Gross et al. 1989) http://course1.winona.edu/kbates/Immunology/images/figur

e_05_06.jpg

Chimeric Antigen Receptor (CAR)

● Step 2: Create single chain variable fragments (scFv’s)○ VH and VL obtain from antibodies

to make scFv

● Step 3: Zeta chain from T-Cell signalling complex attached by spacer.

http://upload.wikimedia.org/wikipedia/commons/4/47/CAR_cartoon.png

● Step 1: Create monoclonal antibodies○ mice or human tumor cells in lab

http://www.biology.arizona.edu/immunology/tutorials/antibody/graphics/antibody.gif

Generations of CARs

http://www.hindawi.com/journals/bmri/2010/956304/fig3/

Effects of CAR

• Recognition of tumor associated antigen (TAA) on target cell via CAR sends activating signal

• Once activated it triggers release of perforin and granzymes along with cytokines by other immune cells

http://www.hindawi.com/journals/bmri/2010/956304/fig2/ http://classes.midlandstech.edu/carterp/Courses/bio225/chap17/Slide34_211.jpg

Delivery Method

◼ Mainly through viral vectors (adenovirus and retrovirus are most common)

◼ Top pictures: adenovirus structure and method of infecting host cell

◼ Bottom pictures: retrovirus structure and method of integration.

→

→

http://www.genetherapynet.com/viral-vectors/adenoviruses.html

http://www.genetherapynet.com/viral-vectors/adenoviruses.html

http://www.ohsu.edu/xd/about/services/integrity/upload/IBC_Presentation-Choosing-a-Viral-Vector-System.pdf http://www.ohsu.edu/xd/about/services/integrity/upload/IBC_Presentation-Choosing-a-Viral-Vector-System.pdf

Correct Vector is Dependent on Event

◼ Type of target cell (dividing or non-dividing)

◼ Short-term or long-term expression◼ Retrovirus:

▪ Long-term expression▪ Infect dividing cells ▪ Integrates into target cell genome

◼ Adenovirus:▪ Can infect dividing and non-dividing cells▪ Short-term gene expression▪ Does not integrate into target cell genome

Patient Involvement

• Isolate WBCs from patient

• Genetically modified T-cells in vitro

• Expand modified T-cells

• Reintroduce patients own modified T-cells with CAR back into circulation

• Monitor and run tests for proliferation in vivo

http://clincancerres.aacrjournals.org/content/18/10/2780/F2.large.jpg

Team at Penn Medicine

• In 2013, team at Penn Medicine was nationally recognized in top newspapers for breakthrough results with CAR immunotherapy study of first 59 adults and children with chronic lymphocytic leukemia or acute lymphoblastic leukemia

• Response to therapy:

• 15 of 32 adults with CLL • 19 of 22 pediatric with ALL• 5 of 5 adults with ALLhttp://www.uphs.upenn.edu/news/News_Releases/

2013/12/ctl019/

References

◼ 1) Friedmann, T., & Roblin, R. (1972). Gene Therapy for Human Genetic Disease? Science, 175(4025), 949-955. Retrieved March 13, 2014, from http://www.jstor.org.ozone.nsc.edu/stable/1732705

◼ 2) Blaese, R. M., Culver, K. W., Miller, A. D., Carter, C. S., Fleisher, T., Clerici, M., ... Anderson, W. F. (1995). T Lymphocyte-Directed Gene Therapy for ADA$^-$ SCID: Initial Trial Results After 4 Years. Science, 270(5235), 475-480. Retrieved March 13, 2014, from http://www.jstor.org/stable/2889066

◼ 3) Abramson Cancer Center. (n.d.). Retrieved March 13, 2014, from https://www.penncancer.org/penn_news.cfm?ID=2125◼ 4) Penn Medicine Team Reports Findings from Research Study of First 59 Adult and Pediatric Leukemia Patients Who

Received Investigational, Personalized Cellular Therapy CTL019. (n.d.). Retrieved March 13, 2014, from http://www.uphs.upenn.edu/news/News_Releases/2013/12/ctl019/

◼ 5) Marchione, M. (2013, December 8). Gene therapy scores big wins against blood cancers. NBC News. Retrieved March 13, 2014, from http://www.nbcnews.com/health/cancer/gene-therapy-scores-big-wins-against-blood-cancers-f2D11708740

◼ 6) Vorburger, S. A., & Hunt, K. K. (2002, February 01). Adenoviral Gene Therapy. The Oncologist, 7(1), 46-59. Retrieved March 13, 2014, from http://theoncologist.alphamedpress.org/content/7/1/46.abstract

◼ 7) Turtle, C. (2013). Chimeric antigen receptor modified T cell therapy for B cell malignancies. International Journal of Hematology, 99(2), 132-140. Retrieved March 13, 2014, from http://link.springer.com.ozone.nsc.edu:8080/article/10.1007%2Fs12185-013-1490-x/fulltext.html

◼ 8) Chekmasova, A. A., & Brentjens, R. J. (2010). Adoptive T Cell Immunotherapy Strategies for the Treatment of Patients with Ovarian Cancer - Alena A Chekmasova - Discovery Medicine. Discovery Medicine. Retrieved March 13, 2014, from http://www.discoverymedicine.com/Alena-A-Chekmasova/2010/01/22/adoptive-t-cell-immunotherapy-strateg....

◼ 9) Lee, D. W., Barrett, D. M., Mackall, C., Orentas, R., & Grupp, S. A. (2012, May 15). The Future Is Now: Chimeric Antigen Receptors as New Targeted Therapies for Childhood Cancer. Clinical Cancer Research, 18(10), 2780-2790. Retrieved March 13, 2014, from http://clincancerres.aacrjournals.org/content/18/10/2780.abstract

◼ 10) Cancer Research Updates. (n.d.). Retrieved March 12, 2014, from http://www.cancer.gov/cancertopics/research-updates/2013/CAR-T-Cells

◼ 11) Parham, P., & Janeway, C. (2009). The immune system. London: Garland Science.