In our lab, we have a plethora of cell lines which includes head and neck cancer cells, lung cancer cells, breast cancer cells, colon cancer cells, liver, kidney, fibroblast, etc. Most of our work focuses on lung cancer and head & neck cancer cells, from handling cell culture to screening and cytotoxicity of compounds (mainly synthesized in our lab), such as MTT Assay and Clonogenicity Assay. We have also made a retroviral stable cell line. In addition, assays such as apoptotic assay, cell cycle analysis, fluorescent microscopy studies for ds DNA breaks were studied.
The radioprotectors and radiosensitizers synthesized in the lab were further studiedto identify resistance/ sensitivity mechanisms in the Xenograft model. In the Xenograft model, we study the conventional cell line xenograft and Patient-Derived Xenograft (PDX) forradiation response studies, head and neck cancer, as well as looking at simultaneous radio and chemotherapy treatment which mimics the clinical setting. PDX models provide a renewable source of cancer cells for 3D model generation.
Furthermore, we establish different organoid/ spheroid development methods and study the effect of radiation alone and radiation along with radio protectors.
As per ICMR Cancer 2020, there is approximately 17.3 Lakhs & 8.8 Lakhs cancer death in India. India has over 1 million new cancer cases every year. Radiation is the most widely used therapy in curative treatment for solid cancerous tumors over a long period. Due to severe radiotherapy’s severe adverse effects, the Radiation Oncologist is looking forward to potent Radioprotectors. Amifostine is the only FDA approved drug to reduce the toxicity of radiation therapy in the setting of cancer treatment, but not for ARS and has several clinically relevant limitations. Despite additional research, no potent radioprotector is approved by FDA for clinical relevance. My laboratory developed DMA (5- {4-methylpiperazin-1-yl}-2-[2’-(3,4-dimethoxyphenyl)-5’-benzimidazolyl], as a small synthetic molecule with increased lipophilicity but reduced cytotoxicity. The present invention relates to dual activation of Akt/NFκB pathway by DMA (5-(4-methylpiperazin-1-yl)-2-[2′-(3,4-dimethoxyphenyl)-5′-benzimidazolyl] benzimidazole) to render radioprotection both in mammalian cells and in Balb/c mice. Further, it selectively protects normal cells overs tumor tissues against lethal total body irradiation (TBI). There was no activation of Akt/NFκB pathway by DMA in response to radiation in tumor tissues. A single DMA dose before TBI protects mice from GI and HP acute radiation syndrome (ARS) and offered radioprotection through oral, i.v., i.p., and s.c route of administration. DMA’s half-life in plasma is 3.5 h at oral dose and 90% clearance was observed in 16 h. DMA accumulates in high concentration in intestine, liver, kidney, and spleen tissues, justifying the observed radioprotection to normal tissue even at a single dose. This data provide a molecular rationale that DMA is a selective radioprotector to normal tissue and can improve radiotherapy’s clinical outcome, valuable as adjuvants in cancer therapy and management of radiation emergencies. Results in Balb/c mice against whole-body irradiation showed radioprotection by DMA when delivered orally 2h before radiation. It showed dose-dependent radioprotection at 100, 150, 200 and 400mg/kg bw doses of DMA at 8 Gy. In addition to the above, a series of small animal experiments showed that there was 100% and 50% mice survival at 8 and 9 Gy TBI respectively by systemic administration of DMA. In contrast, it was 55% by i.p. and 20% through s.c. administration routes. Histopathology of the tissues demonstrate that DMA ameliorate radiation induced damage of hematopoietic and gastrointestinal syndromes. All these data indicate that DMA specifically protects normal cells over tumor cells. Indian Patent, US Patent, and EU patent granted. (For more detail click here)
DMA, A Bisbenzimidazole to take it to Early-Phase Human Phase Trials as a Radio modulator/Radioprotector to Normal Cells for Cancer Radiotherapy in Patients.
Prochlorperazine enhances radiosensitivity of Non-Small Cell Lung Carcinoma by stabilizing GDP-bound mutant KRAS Conformation. The docking studies revealed the H-bond of the pendant piperazine ring -NH with the backbone carbonyl of Tyr-32. MD simulation showcased that PCZ undergoes fluctuations in all variants, and the piperazine group drifts away from the Tyr-32 residue into the solvent. Evidently, the piperazine seems to elicit more favorable interactions with the bulk solvent rather than the protein. However, the tricyclic ring doesn’t move away from the guanine binding site completely except in one case (G12D) during the 50ns simulation. This suggests a robust interaction of PCZ with the guanine binding site, even if it is made up of several small dispersion interactions with the switch-I and other regions of K-Ras. The stabilization of the switch-I region in G12V, G12S, and G12C mutants compared to WT argues in favour of PCZ binding to the GDP-bound conformation of K-Ras. The behaviour of the G12A-PCZ complex serves as a negative control for this simulation experiment as PCZ is not able to stabilize the GTP-bound conformation.