Treating Diffuse Large B Cell Lymphoma Using HLA Class I Molecule Deficient Anti CD19 CAR-NK Cells

Student: Suraj Das
Table: MED10
Experimentation location: Home
Regulated Research (Form 1c): No
Project continuation (Form 7): No

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  1. Marshall, J.S., Warrington, R., Watson, W. et al. An introduction to immunology and immunopathology. Allergy Asthma Clin Immunol 14, 49 (2018).


  1. Chaplin DD. Overview of the immune response. J Allergy Clin Immunol. 2010 Feb;125(2 Suppl 2):S3-23. doi: 10.1016/j.jaci.2009.12.980. PMID: 20176265; PMCID: PMC2923430.


  1. Alberts B, Johnson A, Lewis J, et al. Molecular Biology of the Cell. 4th edition. New York: Garland Science; 2002. Innate Immunity. Available from:


  1. Gasteiger G, D'Osualdo A, Schubert D, A, Weber A, Bruscia E, M, Hartl D: Cellular Innate Immunity: An Old Game with New Players. J Innate Immun 2017;9:111-125. doi: 10.1159/000453397


  1. Alberts B, Johnson A, Lewis J, et al. Molecular Biology of the Cell. 4th edition. New York: Garland Science; 2002. Chapter 24, The Adaptive Immune System. Available from:


  1. Janeway CA Jr, Travers P, Walport M, et al. Immunobiology: The Immune System in Health and Disease. 5th edition. New York: Garland Science; 2001. The components of the immune system. Available from:


  1. Kondo M. Lymphoid and myeloid lineage commitment in multipotent hematopoietic progenitors. Immunol Rev. 2010 Nov;238(1):37-46. doi: 10.1111/j.1600-065X.2010.00963.x. PMID: 20969583; PMCID: PMC2975965.


  1. Cano RLE, Lopera HDE. Introduction to T and B lymphocytes. In: Anaya JM, Shoenfeld Y, Rojas-Villarraga A, et al., editors. Autoimmunity: From Bench to Bedside [Internet]. Bogota (Colombia): El Rosario University Press; 2013 Jul 18. Chapter 5. Available from:


  1. Groscurth P. Cytotoxic effector cells of the immune system. Anat Embryol (Berl). 1989;180(2):109-19. doi: 10.1007/BF00309762. PMID: 2679226.


  1. Paul Sourav, Lal Girdhari. The Molecular Mechanism of Natural Killer Cells Function and Its Importance in Cancer Immunotherapy: Frontiers in Immunology. 8. 2017 10.3389/fimmu.2017.01124  1664-3224   


  1. CLUSTER OF DIFFERENTIATION (CD) ANTIGENS. Immunology Guidebook. 2004:47–124. doi: 10.1016/B978-012198382-6/50027-3. Epub 2007 May 9. PMCID: PMC7158181.


  1. Kumar BV, Connors TJ, Farber DL. Human T Cell Development, Localization, and Function throughout Life. Immunity. 2018 Feb 20;48(2):202-213. doi: 10.1016/j.immuni.2018.01.007. PMID: 29466753; PMCID: PMC5826622.


  1. Wensveen FM, Jelenčić V, Polić B. NKG2D: A Master Regulator of Immune Cell Responsiveness. Front Immunol. 2018 Mar 8;9:441. doi: 10.3389/fimmu.2018.00441. PMID: 29568297; PMCID: PMC5852076.


  1. Sivori, S., Vacca, P., Del Zotto, G. et al. Human NK cells: surface receptors, inhibitory checkpoints, and translational applications. Cell Mol Immunol 16, 430–441 (2019).


  1. Cruz-Tapias P, Castiblanco J, Anaya JM. Major histocompatibility complex: Antigen processing and presentation. In: Anaya JM, Shoenfeld Y, Rojas-Villarraga A, et al., editors. Autoimmunity: From Bench to Bedside [Internet]. Bogota (Colombia): El Rosario University Press; 2013 Jul 18. Chapter 10. Available from:


  1. Wieczorek M, Abualrous ET, Sticht J, Álvaro-Benito M, Stolzenberg S, Noé F, Freund C. Major Histocompatibility Complex (MHC) Class I and MHC Class II Proteins: Conformational Plasticity in Antigen Presentation. Front Immunol. 2017 Mar 17;8:292. doi: 10.3389/fimmu.2017.00292. PMID: 28367149; PMCID: PMC5355494.


  1. Janeway CA Jr, Travers P, Walport M, et al. Immunobiology: The Immune System in Health and Disease. 5th edition. New York: Garland Science; 2001. The major histocompatibility complex and its functions. Available from:


  1. Matasar MJ, Zelenetz AD. Overview of lymphoma diagnosis and management. Radiol Clin North Am. 2008 Mar;46(2):175-98, vii. doi: 10.1016/j.rcl.2008.03.005. PMID: 18619375.


  1. de Leval L, Jaffe ES. Lymphoma Classification. Cancer J. 2020 May/Jun;26(3):176-185. doi: 10.1097/PPO.0000000000000451. PMID: 32496451.


  1. Aggarwal P, Limaiem F. Reed Sternberg Cells. [Updated 2021 Jul 26]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2022 Jan-. Available from:


  1. Roschewski M, Phelan JD, Wilson WH. Molecular Classification and Treatment of Diffuse Large B-Cell Lymphoma and Primary Mediastinal B-Cell Lymphoma. Cancer J. 2020 May/Jun;26(3):195-205. doi: 10.1097/PPO.0000000000000450. PMID: 32496453; PMCID: PMC7285963.


  1. Bakshi N, Maghfoor I. The current lymphoma classification: new concepts and practical applications triumphs and woes. Ann Saudi Med. 2012 May-Jun;32(3):296-305. doi: 10.5144/0256-4947.2012.296. PMID: 22588443; PMCID: PMC6081048.


  1. Nowakowski GS, Czuczman MS. ABC, GCB, and Double-Hit Diffuse Large B-Cell Lymphoma: Does Subtype Make a Difference in Therapy Selection? Am Soc Clin Oncol Educ Book. 2015:e449-57. doi: 10.14694/EdBook_AM.2015.35.e449. PMID: 25993209.


  1. Coiffier B, Sarkozy C. Diffuse large B-cell lymphoma: R-CHOP failure-what to do? Hematology Am Soc Hematol Educ Program. 2016 Dec 2;2016(1):366-378. doi: 10.1182/asheducation-2016.1.366. PMID: 27913503; PMCID: PMC6142522.


  1. Hopfinger G, Jäger U, Worel N. CAR-T Cell Therapy in Diffuse Large B Cell Lymphoma: Hype and Hope. Hemasphere. 2019 Mar 8;3(2):e185. doi: 10.1097/HS9.0000000000000185. PMID: 31723824; PMCID: PMC6746029.


  1. Lu H, Zhao X, Li Z, Hu Y, Wang H. From CAR-T Cells to CAR-NK Cells: A Developing Immunotherapy Method for Hematological Malignancies. Front Oncol. 2021 Aug 6;11:720501. doi: 10.3389/fonc.2021.720501. PMID: 34422667; PMCID: PMC8377427


  1. Albinger, N., Hartmann, J. & Ullrich, E. Current status and perspective of CAR-T and CAR-NK cell therapy trials in Germany. Gene Ther 28, 513–527 (2021).


  1. Hopfinger G, Jäger U, Worel N. CAR-T Cell Therapy in Diffuse Large B Cell Lymphoma: Hype and Hope. Hemasphere. 2019 Mar 8;3(2):e185. doi: 10.1097/HS9.0000000000000185. PMID: 31723824; PMCID: PMC6746029.


  1. David Sermer, Connie Batlevi, M. Lia Palomba, Gunjan Shah, Richard J. Lin, Miguel-Angel Perales, Michael Scordo, Parastoo Dahi, Martina Pennisi, Aishat Afuye, Mari Lynne Silverberg, Caleb Ho, Jessica Flynn, Sean Devlin, Philip Caron, Audrey Hamilton, Paul Hamlin, Steven Horwitz, Erel Joffe, Anita Kumar, Matthew Matasar, Ariela Noy, Colette Owens, Alison Moskowitz, David Straus, Gottfried von Keudell, Ildefonso Rodriguez-Rivera, Lorenzo Falchi, Andrew Zelenetz, Joachim Yahalom, Anas Younes, Craig Sauter; Outcomes in patients with DLBCL treated with commercial CAR T cells compared with alternate therapies. Blood Adv 2020; 4 (19): 4669–4678. doi:


  1. Neelapu SS, Locke FL, Bartlett NL, Lekakis LJ, Miklos DB, Jacobson CA, Braunschweig I, Oluwole OO, Siddiqi T, Lin Y, Timmerman JM, Stiff PJ, Friedberg JW, Flinn IW, Goy A, Hill BT, Smith MR, Deol A, Farooq U, McSweeney P, Munoz J, Avivi I, Castro JE, Westin JR, Chavez JC, Ghobadi A, Komanduri KV, Levy R, Jacobsen ED, Witzig TE, Reagan P, Bot A, Rossi J, Navale L, Jiang Y, Aycock J, Elias M, Chang D, Wiezorek J, Go WY. Axicabtagene Ciloleucel CAR T-Cell Therapy in Refractory Large B-Cell Lymphoma. N Engl J Med. 2017 Dec 28;377(26):2531-2544. doi: 10.1056/NEJMoa1707447. Epub 2017 Dec 10. PMID: 29226797; PMCID: PMC5882485.


  1. Kochenderfer JN, Somerville RPT, Lu T, Yang JC, Sherry RM, Feldman SA, McIntyre L, Bot A, Rossi J, Lam N, Rosenberg SA. Long-Duration Complete Remissions of Diffuse Large B Cell Lymphoma after Anti-CD19 Chimeric Antigen Receptor T Cell Therapy. Mol Ther. 2017 Oct 4;25(10):2245-2253. doi: 10.1016/j.ymthe.2017.07.004. Epub 2017 Jul 13. PMID: 28803861; PMCID: PMC5628864.


  1. Adli, M. The CRISPR tool kit for genome editing and beyond. Nat Commun 9, 1911 (2018).


  1. Loureiro A, da Silva GJ. CRISPR-Cas: Converting A Bacterial Defense Mechanism into A State-of-the-Art Genetic Manipulation Tool. Antibiotics (Basel). 2019 Feb 28;8(1):18. doi: 10.3390/antibiotics8010018. PMID: 30823430; PMCID: PMC6466564.


  1. Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero DA, Horvath P. CRISPR provides acquired resistance against viruses in prokaryotes. Science. 2007 Mar 23;315(5819):1709-12. doi: 10.1126/science.1138140. PMID: 17379808.


  1. Shabbir, M.A.B., Shabbir, M.Z., Wu, Q. et al. CRISPR-cas system: biological function in microbes and its use to treat antimicrobial resistant pathogens. Ann Clin Microbiol Antimicrob 18, 21 (2019).


  1. Nidhi S, Anand U, Oleksak P, Tripathi P, Lal JA, Thomas G, Kuca K, Tripathi V. Novel CRISPR-Cas Systems: An Updated Review of the Current Achievements, Applications, and Future Research Perspectives. Int J Mol Sci. 2021 Mar 24;22(7):3327. doi: 10.3390/ijms22073327. PMID: 33805113; PMCID: PMC8036902.


  1. Rautela I, Uniyal P, Thapliyal P, Chauhan N, Bhushan Sinha V, Dev Sharma M. An extensive review to facilitate understanding of CRISPR technology as a gene editing possibility for enhanced therapeutic applications. Gene. 2021 Jun 15;785:145615. doi: 10.1016/j.gene.2021.145615. Epub 2021 Mar 26. PMID: 33775851.


  1. Allen D, Rosenberg M, Hendel A. Using Synthetically Engineered Guide RNAs to Enhance CRISPR Genome Editing Systems in Mammalian Cells. Front Genome Ed. 2021 Jan 28;2:617910. doi: 10.3389/fgeed.2020.617910. PMID: 34713240; PMCID: PMC8525374.


  1. Cui, Y., Xu, J., Cheng, M. et al. Review of CRISPR/Cas9 sgRNA Design Tools. Interdiscip Sci Comput Life Sci 10, 455–465 (2018).


  1. Gleditzsch D, Pausch P, Müller-Esparza H, Özcan A, Guo X, Bange G, Randau L. PAM identification by CRISPR-Cas effector complexes: diversified mechanisms and structures. RNA Biol. 2019 Apr;16(4):504-517. doi: 10.1080/15476286.2018.1504546. Epub 2018 Sep 18. PMID: 30109815; PMCID: PMC6546366.


  1. Orii, Kenji E., et al. “Selective Utilization of Nonhomologous End-Joining and Homologous Recombination DNA Repair Pathways during Nervous System Development.” Proceedings of the National Academy of Sciences of the United States of America, vol. 103, no. 26, National Academy of Sciences, 2006, pp. 10017–22,


  1. Gong, Y., Klein Wolterink, R.G.J., Wang, J. et al. Chimeric antigen receptor natural killer (CAR-NK) cell design and engineering for cancer therapy. J Hematol Oncol 14, 73 (2021).


  1. Marofi, F., Saleh, M.M., Rahman, H.S. et al. CAR-engineered NK cells; a promising therapeutic option for treatment of hematological malignancies. Stem Cell Res Ther 12, 374 (2021).


  1. Khantasup K, Chantima W, Sangma C, Poomputsa K, Dharakul T. Design and Generation of Humanized Single-chain Fv Derived from Mouse Hybridoma for Potential Targeting Application. Monoclon Antib Immunodiagn Immunother. 2015 Dec;34(6):404-17. doi: 10.1089/mab.2015.0036. PMID: 26683180; PMCID: PMC4685505.


  1. Fujiwara K, Masutani M, Tachibana M, Okada N. Impact of scFv structure in chimeric antigen receptor on receptor expression efficiency and antigen recognition properties. Biochem Biophys Res Commun. 2020 Jun 25;527(2):350-357. doi: 10.1016/j.bbrc.2020.03.071. Epub 2020 Mar 23. PMID: 32216966.


  1. Thokala R, Olivares S, Mi T, Maiti S, Deniger D, Huls H, Torikai H, Singh H, Champlin RE, Laskowski T, McNamara G, Cooper LJ. Redirecting Specificity of T cells Using the Sleeping Beauty System to Express Chimeric Antigen Receptors by Mix-and-Matching of VL and VH Domains Targeting CD123+ Tumors. PLoS One. 2016 Aug 22;11(8):e0159477. doi: 10.1371/journal.pone.0159477. PMID: 27548616; PMCID: PMC4993583.


  1. Ying, Z., Huang, X.F., Xiang, X. et al. A safe and potent anti-CD19 CAR T cell therapy. Nat Med 25, 947–953 (2019).


  1. Alabanza L, Pegues M, Geldres C, Shi V, Wiltzius JJW, Sievers SA, Yang S, Kochenderfer JN. Function of Novel Anti-CD19 Chimeric Antigen Receptors with Human Variable Regions Is Affected by Hinge and Transmembrane Domains. Mol Ther. 2017 Nov 1;25(11):2452-2465. doi: 10.1016/j.ymthe.2017.07.013. Epub 2017 Jul 27. PMID: 28807568; PMCID: PMC5675490.


  1. Li Y, Hermanson DL, Moriarity BS, Kaufman DS. Human iPSC-Derived Natural Killer Cells Engineered with Chimeric Antigen Receptors Enhance Anti-tumor Activity. Cell Stem Cell. 2018 Aug 2;23(2):181-192.e5. doi: 10.1016/j.stem.2018.06.002. Epub 2018 Jun 28. PMID: 30082067; PMCID: PMC6084450.


  1. Salter AI, Ivey RG, Kennedy JJ, Voillet V, Rajan A, Alderman EJ, Voytovich UJ, Lin C, Sommermeyer D, Liu L, Whiteaker JR, Gottardo R, Paulovich AG, Riddell SR. Phosphoproteomic analysis of chimeric antigen receptor signaling reveals kinetic and quantitative differences that affect cell function. Sci Signal. 2018 Aug 21;11(544):eaat6753. doi: 10.1126/scisignal.aat6753. PMID: 30131370; PMCID: PMC6186424.


  1. Gargett T, Brown MP. The inducible caspase-9 suicide gene system as a "safety switch" to limit on-target, off-tumor toxicities of chimeric antigen receptor T cells. Front Pharmacol. 2014 Oct 28;5:235. doi: 10.3389/fphar.2014.00235. PMID: 25389405; PMCID: PMC4211380.


  1. Chmielewski M, Abken H. TRUCKs: the fourth generation of CARs. Expert Opin Biol Ther. 2015;15(8):1145-54. doi: 10.1517/14712598.2015.1046430. Epub 2015 May 18. PMID: 25985798.


  1. Klingemann H, Boissel L, Toneguzzo F. Natural Killer Cells for Immunotherapy - Advantages of the NK-92 Cell Line over Blood NK Cells. Front Immunol. 2016 Mar 14;7:91. doi: 10.3389/fimmu.2016.00091. PMID: 27014270; PMCID: PMC4789404.


  1. Klingemann HP33. NK-92 cellular immunotherapy as an alternative to donor derived peripheral blood NK cellsJournal for ImmunoTherapy of Cancer 2014;2:P24. doi: 10.1186/2051-1426-2-S2-P24


  1. David A. Knorr, Zhenya Ni, David Hermanson, Melinda K. Hexum, Laura Bendzick, Laurence J.N. Cooper, Dean A. Lee, Dan S. Kaufman, Clinical-Scale Derivation of Natural Killer Cells From Human Pluripotent Stem Cells for Cancer Therapy, Stem Cells Translational Medicine, Volume 2, Issue 4, April 2013, Pages 274–283,


  1. Ni Z, Knorr DA, Clouser CL, Hexum MK, Southern P, Mansky LM, Park IH, Kaufman DS. Human pluripotent stem cells produce natural killer cells that mediate anti-HIV-1 activity by utilizing diverse cellular mechanisms. J Virol. 2011 Jan;85(1):43-50. doi: 10.1128/JVI.01774-10. Epub 2010 Oct 20. PMID: 20962093; PMCID: PMC3014194.


  1. Stefanie Raab, Moritz Klingenstein, Stefan Liebau, Leonhard Linta, "A Comparative View on Human Somatic Cell Sources for iPSC Generation", Stem Cells International, vol. 2014, Article ID 768391, 12 pages, 2014.


  1. Itoh M, Kiuru M, Cairo MS, Christiano AM. Generation of keratinocytes from normal and recessive dystrophic epidermolysis bullosa-induced pluripotent stem cells. Proc Natl Acad Sci U S A. 2011 May 24;108(21):8797-802. doi: 10.1073/pnas.1100332108. Epub 2011 May 9. PMID: 21555586; PMCID: PMC3102348.


  1. Marofi F, Al-Awad AS, Sulaiman Rahman H, Markov A, Abdelbasset WK, Ivanovna Enina Y, Mahmoodi M, Hassanzadeh A, Yazdanifar M, Stanley Chartrand M, Jarahian M. CAR-NK Cell: A New Paradigm in Tumor Immunotherapy. Front Oncol. 2021 Jun 10;11:673276. doi: 10.3389/fonc.2021.673276. PMID: 34178661; PMCID: PMC8223062.


  1. Aasen, T., Raya, A., Barrero, M. et al. Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes. Nat Biotechnol 26, 1276–1284 (2008).


  1. Giorgetti A, Montserrat N, Aasen T, Gonzalez F, Rodríguez-Pizà I, Vassena R, Raya A, Boué S, Barrero MJ, Corbella BA, Torrabadella M, Veiga A, Izpisua Belmonte JC. Generation of induced pluripotent stem cells from human cord blood using OCT4 and SOX2. Cell Stem Cell. 2009 Oct 2;5(4):353-7. doi: 10.1016/j.stem.2009.09.008. PMID: 19796614; PMCID: PMC2779776.


  1. Eguizabal C, Zenarruzabeitia O, Monge J, Santos S, Vesga MA, Maruri N, Arrieta A, Riñón M, Tamayo-Orbegozo E, Amo L, Larrucea S, Borrego F. Natural killer cells for cancer immunotherapy: pluripotent stem cells-derived NK cells as an immunotherapeutic perspective. Front Immunol. 2014 Sep 15;5:439. doi: 10.3389/fimmu.2014.00439. PMID: 25309538; PMCID: PMC4164009.


  1. Knorr DA, Ni Z, Hermanson D, Hexum MK, Bendzick L, Cooper LJ, Lee DA, Kaufman DS. Clinical-scale derivation of natural killer cells from human pluripotent stem cells for cancer therapy. Stem Cells Transl Med. 2013 Apr;2(4):274-83. doi: 10.5966/sctm.2012-0084. Epub 2013 Mar 20. PMID: 23515118; PMCID: PMC3659832.


  1. Hoerster K, Uhrberg M, Wiek C, Horn PA, Hanenberg H, Heinrichs S. HLA Class I Knockout Converts Allogeneic Primary NK Cells Into Suitable Effectors for "Off-the-Shelf" Immunotherapy. Front Immunol. 2021 Jan 29;11:586168. doi: 10.3389/fimmu.2020.586168. PMID: 33584651; PMCID: PMC7878547.


  1. Crew MD, Cannon MJ, Phanavanh B, Garcia-Borges CN. An HLA-E single chain trimer inhibits human NK cell reactivity towards porcine cells. Mol Immunol. 2005 Jun;42(10):1205-14. doi: 10.1016/j.molimm.2004.11.013. Epub 2005 Jan 8. PMID: 15829309.


  1. Kang CH, Kim Y, Lee HK, Lee SM, Jeong HG, Choi SU, Park CH. Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line. Int J Mol Sci. 2020 Dec 1;21(23):9163. doi: 10.3390/ijms21239163. PMID: 33271901; PMCID: PMC7730610.


  1. Weinkove R, George P, Dasyam N, McLellan AD. Selecting costimulatory domains for chimeric antigen receptors: functional and clinical considerations. Clin Transl Immunology. 2019 May 11;8(5):e1049. doi: 10.1002/cti2.1049. PMID: 31110702; PMCID: PMC6511336.


  1. Guedan S, Calderon H, Posey AD Jr, Maus MV. Engineering and Design of Chimeric Antigen Receptors. Mol Ther Methods Clin Dev. 2018 Dec 31;12:145-156. doi: 10.1016/j.omtm.2018.12.009. PMID: 30666307; PMCID: PMC6330382.


  1. Tumor cell-intrinsic PD-1 receptor is a tumor suppressor and mediates resistance to PD-1 blockade therapy


  1. Xiaodong Wang, Xiaohui Yang, Chang Zhang, Yang Wang, Tianyou Cheng, Liqiang Duan, Zhou Tong, Shuguang Tan, Hangjie Zhang, Phei Er Saw, Yinmin Gu, Jinhua Wang, Yibi Zhang, Lina Shang, Yajuan Liu, Siyuan Jiang, Bingxue Yan, Rong Li, Yue Yang, Jie Yu, Yunzhao Chen, George Fu Gao, Qinong Ye, Shan Gao: Proceedings of the National Academy of Sciences Mar 2020, 117 (12) 6640-6650; DOI: 10.1073/pnas.1921445117


  1. Han Y, Liu D, Li L. PD-1/PD-L1 pathway: current researches in cancer. Am J Cancer Res. 2020 Mar 1;10(3):727-742. PMID: 32266087; PMCID: PMC7136921.


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  1. Xie G, Dong H, Liang Y, Ham JD, Rizwan R, Chen J. CAR-NK cells: A promising cellular immunotherapy for cancer. EBioMedicine. 2020 Sep;59:102975. doi: 10.1016/j.ebiom.2020.102975. Epub 2020 Aug 24. PMID: 32853984; PMCID: PMC7452675.










































































Additional Project Information

Project website: -- No project website --
Additional Resources: -- No resources provided --

Research Plan:


  • This project is completely research-based, and all of my experimental evidence comes from similar experiments conducted by various labs. Below is a rough outline of my paper, which describes all the topics I did research on to develop my treatment
    • Introduction
      • The immune system
        • Adaptive Immunity
          • T Cells
          • B Cells
        • Innate Immunity
          • Nk Cells
        • Cluster of Differentiation
      • Major Histocompatibility Complex
        • MHC mismatch regulation
      • Chimeric Antigen Receptors
        • CAR Receptor design and structure
        • Types of CAR-cells
        • Shortcomings of CAR cells
      • Lymphoma
        • Non-Hodgkin’s Lymphoma 
          • Diffuse Large B-cell Lymphoma
            • Current treatments
              • Shortcomings
              • Advantages
        • Lymphoma classification


  • CRISPR Cas9 Gene Editing Technology
    • Type II CRISPR systems
    • Previous ways CRISPR has been used in the medical field


  • Nk Cell Sources
    • Induced Pluripotent Stem Cells
    • Human Pluripotent Stem Cells
  • Cell Design
    • Advantageous genes to inhibit function of in NK cells
      • Pd-1 inhibiting receptor
    • CAR-receptor structure to use
  • Experimental Design


  • Functional Assays


  • Conclusion






Questions and Answers

1. What was the major objective of your project and what was your plan to achieve it? 

I desired to make a possible treatment for Diffuse Large B-cell lymphoma that would be capable of overcoming some of the disadvantages current treatments for the disease have.

       a. Was that goal the result of any specific situation, experience, or problem you encountered?  

This goal was not the result of any situation I personally experienced, however, it was the result of an accumulation of experiences in varous fields of science that made me more and more interested in the entire field of medical treatment and how CRISPR Cas9 can be used to treat different cancers.



       b. Were you trying to solve a problem, answer a question, or test a hypothesis?

I was trying to develop a hypothetical solution to my hypothesis. Unfortunately, because of the funding that is required to actually conduct such an experiment properly, it is not feasible to test my hypothesis. I can only use data from already conducted experiments to create my own solution.


2. What were the major tasks you had to perform in order to complete your project?

      There were three major tasks that I needed to complete - background research, experiment, and cell design, and my conclusion. My background research comprised of all the biological topics related to CRISPR Cas9, the immune system, and lymphoma. For further information, see my research plan. My experiment and cell design consisted of my hypothetical CAR-Nk cell structure, as well as the process for creating it.


3. What is new or novel about your project?

       a. My project is entirely unique in the fact that it integrates cell designs from other various experiments, as well as using some original and novel cell designs proposed by me. Primarily, I have conducted two rounds of CRISPR Cas9 gene editing to knock out two different genes that are highly related to Diffuse Large B-cell Lymphoma(DLBCL). Most other treatments have only used one round of gene editing, however in my case I chose to use two because I believe that using such a cell design will increase the Nk cell's targeting capability of malignant B-cells, and also its ability to bypass the HLA mismatch restriction.


4. What was the most challenging part of completing your project?

      The most challenging part of my project was completing the cell design and experimental method, simply because it is very difficult to decide where to start; there is such a vast amount of information regarding my topic. My treatment also had to be something that was overcoming some of the flaws in modern-day treatments, so that added to the difficulty level somewhat.


5. If you were going to do this project again, are there any things you would you do differently the next time?

If there is something I would have liked to do differently, it would probably be to have the opportunity to observe CRISPR Cas9 gene-editing technology first-hand. This is something that I was not able to do, and I feel as though having such an experience would definitely be helpful for modifying my perspective on CRISPR Cas9 and CAR therapies.


6. Did working on this project give you any ideas for other projects? 

This project also created a new possible research topic for me to explore, which is the creation of Induced Pluripotent Stem Cells. The research I did on them for this project made me realize that there is still much work to do in the field to determine the optimal source of stem cells, and I am quite interested in some of the different ways we can develop induced pluripotent stem cells. An example of this would be keratinocyte-based stem cells, which is a source for stem cells that have not been researched extensively but seems to have incredible potential in the medical field.


7. How did COVID-19 affect the completion of your project?

COVID-19 did not really affect the completion of my project, because the large majority of my project was completed through online research, so I did not struggle too much with my project, fortunately.