Engineering of Anti-CD133 Trispecific Molecule Capable of Inducing NK Expansion and Driving Antibody-Dependent Cell-Mediated Cytotoxicity / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The selective elimination of cancer stem cells (CSCs) in tumor patients is a crucial goal because CSCs cause drug refractory relapse. To improve the current conventional bispecific immune-engager platform, a 16133 bispecific natural killer (NK) cell engager (BiKE), consisting of scFvs binding FcγRIII (CD16) on NK cells and CD133 on carcinoma cells, was first synthesized and a modified interleukin (IL)-15 crosslinker capable of stimulating NK effector cells was introduced. MATERIALS AND METHODS: DNA shuffling and ligation techniques were used to assemble and synthesize the 1615133 trispecific NK cell engager (TriKE). The construct was tested for its specificity using flow cytometry, cytotoxic determinations using chromium release assays, and lytic degranulation. IL-15–mediated expansion was measured using flow-based proliferation assays. The level of interferon (IFN)-γ release was measured because of its importance in the anti-cancer response. RESULTS: 1615133 TriKE induced NK cell–mediated cytotoxicity and NK expansion far greater than that achieved with BiKE devoid of IL-15. The drug binding and induction of cytotoxic degranulation was CD133+ specific and the anti-cancer activity was improved by integrating the IL-15 cross linker. The NK cell–related cytokine release measured by IFN-γ detection was higher than that of BiKE. NK cytokine release studies showed that although the IFN-γ levels were elevated, they did not approach the levels achieved with IL-12/IL-18, indicating that release was not at the supraphysiologic level. CONCLUSION: 1615133 TriKE enhances the NK cell anti-cancer activity and provides a self-sustaining mechanism via IL-15 signaling. By improving the NK cell performance, the new TriKE represents a highly active drug against drug refractory relapse mediated by CSCs.

[Characterization of molecular and biochemical properties of some metastatic breast cancer cells]

Study of the molecular and biochemical properties of gene expression, of the certain breast cancer cell lines: metastatic [e.g. HTB-30 SK-BR-3 and HTB-21 CAMA-1]; non-metastatic [e.g. CRL-2315 HCC70 and HTB-126 HS578T]; estrogen receptor positive [ER[+]; e.g. HTB-22 MCF7 and CRL-2329 HCC1500] and estrogen receptor negative [ER-; CRL2335 HCC1806, HTB-132 MB468] was carried out. After all the breast cancer cell lines, in an appropriate cell propagation medium, have been cultured until enough cells were generated [as controls], and treated with one of the fatty acids omega-3 or omega-6 [as samples of treated cells], cancer cells were isolated, then mRNA of these cells were extracted. Microarray technique was applied for studying the genetic sites of the breast cancer cells, and detection the sites where the fatty acids are affected. Confirmation of the respective results have been undertaken using Real Time Polymerase chain Reaction [Real Time PCR] [RT-PCR]. Data resulted from microarray analysis showed many genes that are involved in signaling, lymphocyte activation, apoptosis, cellular growth and proliferation. Also, the results detect an obvious difference between effect of the two studied fatty acids. Omega-3 [Eicosapentaenoic acid "EPA"] works as up regulator of the ER-breast cancer cell lines, while the omega-6 [arachidonic acid "AA"] works as up regulator of the ER+ breast cancer cell lines. This study explored the importance of the ER status in how breast cancer cells respond to omega-3 and omega-6 fatty acids, and the identification of key biomarkers that may shed light on the differential effects of these fatty acids on breast cancer cell lines. This research collaboration has set the stage for future studies to correlate genetic modification of the ER with the cell response to fatty acids, which may have a future applications in the therapeutic managements