BackgroundPhospholipases A2 (PLA2s) are abundant components of snake venoms that have been extensively studied due to their pharmacological and pathophysiological effects on living organisms. This study aimed to assess the antitumor potential of BthTX-I, a basic myotoxic PLA2 isolated from Bothrops jararacussu venom, by evaluating in vitro processes of cytotoxicity, modulation of the cell cycle and induction of apoptosis in human (HL-60 and HepG2) and murine (PC-12 and B16F10) tumor cell lines.MethodsThe cytotoxic effects of BthTX-I were evaluated on the tumor cell lines HL-60 (promyelocytic leukemia), HepG2 (human hepatocellular carcinoma), PC-12 (murine pheochromocytoma) and B16F10 (murine melanoma) using the MTT method. Flow cytometry technique was used for the analysis of cell cycle alterations and death mechanisms (apoptosis and/or necrosis) induced in tumor cells after treatment with BthTX-I.ResultsIt was observed that BthTX-I was cytotoxic to all evaluated tumor cell lines, reducing their viability in 40 to 50 %. The myotoxin showed modulating effects on the cell cycle of PC-12 and B16F10 cells, promoting delay in the G0/G1 phase. Additionally, flow cytometry analysis indicated cell death mainly by apoptosis. B16F10 was more susceptible to the effects of BthTX-I, with ~40 % of the cells analyzed in apoptosis, followed by HepG2 (~35 %), PC-12 (~25 %) and HL-60 (~4 %).ConclusionsThese results suggest that BthTX-I presents antitumor properties that may be useful for developing new therapeutic strategies against cancer.

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Manufacturing of customized gene or cell therapy products  such as CAR-T cells is complex and depends on release tests and exams that can attest to a consistent quality standard for each product. The quality of CAR-T cell products is subject to donor variation, but also includes the manufacturing environment, as well as the quality and availability of materials and reagents. Quality must be carefully monitored and integrated into the manufacturing process.

Cell therapy with T cells expressing chimeric antigen receptor (CAR-T) is a type of immunotherapy that involves the manipulation and reprogramming of immune cells (T lymphocytes) in order to recognize and kill tumor cells. For use in patients, CAR-T cells must be manufcatured inside a GMP facility according to a established procedure.

The therapy with genetically modified T cells to express chimeric antigen receptors (CAR) is a promising strategy for immunotherapy against cancer. CAR-T cells can specifically recognize antigens on the surface of tumor cells and then effectively kill those cells. Several researchers have presented the development of CAR-T cells for various hematological targets and the treatment of solid tumors. Quality control and preclinical evaluation of these products are essential to demonstrate their safety and efficacy and allow development to the clinical trial phase. This chapter will present relevant guidelines regarding pre-clinical research of CAR-T cell products. Preclinical research on cell therapy products should include in vitro and in vivo pharmacodynamics studies (antitumor activity), pharmacokinetics (proliferation, distribution, and persistence of CAR-T cells in vivo), and animal safety studies.