Evaluating the elimination of iCasp9-IL15 CAR-T cells using photosensitizers AP20187 in vitro
Nội dung chính của bài viết
Tóm tắt
This study was conducted to evaluate the ability to eliminate iCasp9-IL15 Chimeric Antigen Receptor-T (CAR-T) cells using the photosensitizers AP20187 in vitro. Peripheral blood mononuclear cells (PBMC) were activated with Dynabeads Human T-Activator CD3/CD28 and IL-2 to optimize the concentration of the antibiotic blasticidin for screening iCasp9-IL15 CAR-T cells. iCasp9-IL15 CAR-T cells after proliferation with 1D2 artificial antigen-presenting cells, were screened using blasticidin. Subsequently, the iCasp9 suicide gene of these cells was activated at various concentrations of AP20187 photosensitizer. The results showed that after 5 days of culture, a blasticidin concentration of 15.0 µg/mL was used to screen iCasp9-IL15 CAR-T cells (activated, non-transformed PBMC showed > 95% cell death). The proportion of iCasp9-IL15 cells after blasticidin screening reached approximately 75%. Using AP20187 at the concentration of 20 nM resulted in almost complete elimination of iCasp9-IL15 CAR-T cells (survival rate: 2.14%). So, iCasp9-IL15 CAR-T cells can almost be eliminated by AP20187 at a concentration of 20 nM in case of high toxicity during treatment.
Chi tiết bài viết
Từ khóa
iCasp9-IL15 CAR-T cells, AP20187 photosensitizer, blasticidin
Tài liệu tham khảo
2. June CH, Sadelain M. Chimeric antigen receptor therapy. New England Journal of Medicine. 2018;379(1):64-73.
3. Bangolo A, Amoozgar B, Mansour C, et al. Comprehensive Review of Early and Late Toxicities in CAR T-Cell Therapy and Bispecific Antibody Treatments for Hematologic Malignancies. Cancers (Basel). 2025;17(2):282.
4. Teachey DT, Lacey SF, Shaw PA, et al. Identification of predictive biomarkers for cytokine release syndrome after chimeric antigen receptor T-cell therapy for acute lymphoblastic leukemia. Cancer Discov. 2016;6(6):664-679.
5. Hoyos V, Savoldo B, Quintarelli C, et al. Engineering CD19-specific T lymphocytes with interleukin-15 and a suicide gene to enhance their anti-lymphoma/leukemia effects and safety. Leukemia. 2010;24(6):1160-1170.
6. Bento FM, Takeshita D, Sacramento CB, et al. Over expression of the selectable marker blasticidin S deaminase gene is toxic to human keratinocytes and murine BALB/MK cells. BMC Biotechnol. 2004;4:1-10.
7. Buck JM, Bártulos CR, Gruber A, et al. Blasticidin-S deaminase, a new selection marker for genetic transformation of the diatom Phaeodactylum tricornutum. PeerJ. 2018;6:e5884.
8. Asoshina M, Myo G, Tada N, et al. Targeted amplification of a sequence of interest in artificial chromosome in mammalian cells. Nucleic Acids Res. 2019;47(11):5998-6006.
9. Hanswillemenke A, Stafforst T. Protocols for the generation of caged guideRNAs for light-triggered RNA-targeting with SNAP-ADARs. Methods in Enzymology. Elsevier; 2019:47-68.
10. Zhou X, Di Stasi A, Brenner MKJGToSCM. iCaspase 9 suicide gene system. Methods Mol Biol. 2015:87-105.
11. Gargett T, Brown MPJFip. 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;5:235.
12. Yu S, Yi M, Qin S, et al. Next generation chimeric antigen receptor T cells: safety strategies to overcome toxicity. Mol Cancer. 2019;18:1-13.
13. Liu Z, Chen O, Wall JBJ, et al. Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector. Sci Rep. 2017;7(1):2193.