Day 12 :
- Cancer Chemotherapy | Cancer: Lifestyle Connection and Nutrition | Cancer Prevention and Research | Cancer Nanotechnology
Location: Zurich, Switzerland
Session Introduction
Gacche Raju
Savitribai Phule Pune University, India
Title: Dietary flavonoid modulates epigenetic markers and miRNA in triple negative breast cancer
Biography:
Rajesh N. Gacche is a Senior professor in the Department of Biotechnology, Savitribai Phule Pune University, India. Prof. Rajesh is a Fellow of Royal Society of Medicine, London, UK. His research focuses on cancer biology, especially exploring natural products and promising plant-derived lead fractions as adjuvants or complementary and alternative medicine for improving the efficacy of conventional drugs and amelioration of emerging drug resistance and off-target toxicities. Prof. Rajesh has published over 150 research papers in reputed international journals. Prof. Rajesh is a author of a book titled as ‘Dietary Research and Cancer (1st ,1). Singapore: Springer Nature 2022. ISBN: 9789811660498. Oncologist, health care & food industry, nursing is widely using this book and the word media has taken cognizance of this book in the mainstream of diet and cancer research.
Abstract:
Statement of the Problem: Triple-negative breast cancer (TNBC) is a heterogeneous group of breast carcinomas distinguished by rapid metastatic growth and aggressive tumor invasion ability, leading to high female mortality. On the eve of emerging drug resistance, there is a need to identify candidate drugs that will improve the efficacy of conventional therapeutic regimes. The purpose of this study was to investigate the effect of dietary flavonoids on regulation of epigenetic markers and miRNAs involved in progression of TNBC.
Methodology & Theoretical Orientation: Series of in vitro cell culture assays such as MTT assay for cell viability, cell migration assay, ROS assay, cell cycle inhibition and apoptosis assay was carried out FACs analysis. The gene expression profile of HDAC 1-11 isomers, DNMT-1/3 and HMT, Onco- and tumor suppressor miRNAs and MRP1 & BCRP was determined using RT-qPCR. The protein expression analysis of HDACs 1, 3, 4 & 6 and apoptosis marker proteins was analyzed using immunoblotting.
Findings: The Apigenin, Galangin and luteolin, showed good potential to modulate the expression profiles of epigenetic regulators in TNBC (MD-MB-231) cells. The HDAC1, HDAC2, HDAC3, HDAC4, HDAC5, HDAC6 and HDAC8 were downregulated in MD-MB-231 cells after treatment with dietary flavonoids such as Apigenin, Galangin and Luteolin. The results were normalized with GAPDH. The protein expression level of HDAC1, HDAC3, HDAC4 and HDAC6 were reduced after the treatment of Apigenin, Galangin and Luteolin. The expression of DNMT1 and HMT downregulated after the treatment of Apigenin, Galangin and Luteolin. The oncomiR-21 was downregulated after the treatment of Apigenin and Luteolin. Whereas, the tumor suppressor miRNAs such as miR34 and miR200b was significantly upregulated after the treatment of Apigenin, Galangin and Luteolin. The combinatorial studies suggested the synergistic effect (CI<1) between flavonoids and SAHA against MDA-MB-231 cells. The qRT-PCR and Western analysis results were supported by molecular docking and MD simulation studies.
Conclusion & Significance: On the eve of evolving drug resistance, tumor heterogeneity, and off target toxicities there is need to find alternative candidate adjuvant therapeutic molecules which will augment the efficacy of conventional drugs and circumvent the drug resistance. The results of the present study clearly indicate the significance of lead flavonoid molecules as adjuvant drug molecules for effective treatment of TNBC.
Recent Publications:
- Choudhari J, Nimma R, Nimal SK, Totakura Venkata SK, Kundu GC, Gacche RN. (2023) Prosopis juliflora (Sw.) DC phytochemicals induce apoptosis and inhibit cell proliferation signaling pathways, EMT, migration, invasion, angiogenesis and stem cell markers in melanoma cell lines. J Ethnopharmacol. 10; 312:116472. (IF 5.2).
- Pote MS, Gacche RN. (2023). ATP-binding cassette efflux transporters and MDR in cancer. Drug Discov Today. 2023 Feb 16:103537. doi: 10.1016/j.drudis.2023.103537. (IF 8.4).
- Deepshikha Singh, Yehuda G. Assaraf, Rajesh N. Gacche (2022). Long Non-coding RNA Mediated Drug Resistance in Breast Cancer. Drug Resistance Updates. 100851, https://doi.org/10.1016/j.drup.2022.10085. (IF 22.84).
- Sonali S. Kamble, Kapil K. Patil, Shrikant V. Hese, Bhaskar S. Dawane, Choudhari Jasoda, Kumar TVS, Rajesh N. Gacche (2022). “Chloroxylon swietenia (Roxb.) DC induces cell death & apoptosis by down-regulating the NF-κB pathway in MCF-7 breast cancer cells: in vitro and in vivo investigations” Cancer Reports e1600.
- Navanath Kumbhar, Snehal Nimal, Sagar Barale, Subodh Kamble, Rohit Bavi, Kailas Sonawane, Rajesh Gacche (2022). Identification of novel leads as potent inhibitors of HDAC3 using ligand-based pharmacophore modeling and MD simulation. Scientific Reports 12(1):1712 (Nature Publication).
Ankesh kumar jaiswal
Indian Institute of Technology Bombay, India
Title: Prior chemotherapy negatively impacts the manufacturing and clinical response of Chimeric Antigen Receptor (CAR) T cell therapeutics in DLBCL: A phase I clinical trial experience from India
Time : 10:30-11:00
Biography:
Ankesh Kumar Jaiswal, have immense passion in the research and development for improving the health and life of cancer patients. He has earned his expertise in the cell and gene therapy specially CAR-T cell therapy. He have worked on the end-to-end work flow of CAR-T cell therapy, starting from construct design to CAR-T cell manufacturing and its functional validation using multiple tools and technique.
Abstract:
Despite remarkable success of CAR-T therapy, complexity of CAR-T manufacturing and high costs limits their accessibility in low-resourced countries including India. Currently, approved CAR-T cells therapy is available to relapsed and refractory patients only. Impact of prior chemotherapy of CAR-T cells manufacturing and clinical response is still a gray area. We have demonstrated efficacy of novel Humanized anti-CD19 CAR-T (HCAR19) cell in preclinical model. Here, we report our experience on scaled manufacturing of HCAR19 from manufacturing to first-in-human Phase I clinical study in r/r B cell malignancies in India along with the impact of prior chemotherapy on patients’ T cells, CAR-T manufacturing and clinical response. To examine the feasibility and functionality of the developed manufacturing process and platform, the end-to-end CAR-T cell manufactured in close system for 10 patients was performed. CD3+ T cells from 10 patients of B cell malignancies were enriched/activated and transduction with anti-CD19 lentivirus vectors. CAR-T were manufactured from all the patient’s T cell successfully (100% success rate) within 8 days of culture (Robust expansion-20-30 fold, TE:>25%, dosage range: 1.78x106-17x106 viable HCAR19/kg) maintaining antitumor activity with memory phenotype. CAR-T product also qualified release criteria for safety, identity, purity and potency. Patients enrolled for clinical trial were segregated into heavily or less treated based on number of prior chemotherapies. Impact of chemotherapy on CAR T cell therapy was measured on parameter of patient’s T cell health, CAR T cells manufacturing capabilities and clinical efficacy. Interestingly, Patients with one line of chemotherapy therapy had superior beneficial impact on patient’s T cells health and CAR-T cell manufacturing benefits while maintaining higher memory cells compared to patients with >/= 1 lines of prior therapies. We developed a novel indigenous anti-CD19 CAR-T product and highly affordable CAR-T manufacturing platform & establish impact of chemotherapy on CAR-T cells manufacturing and its clinical response.