School of Biotechnology (Oct – Dec 2022)

On Jan 5, 2023

JOURNAL ARTICLES

1. Pradhan, R., Paul, S., Das, B., Sinha, S., Dash, S. R., Mandal, M., & Kundu, C. N. (2022). Resveratrol Nanoparticle attenuates metastasis and angiogenesis by deregulating inflammatory cytokines through inhibition of CAFs in oral cancer by CXCL-12/IL-6 dependent pathway. The Journal of Nutritional Biochemistry, 109257. https://doi.org/10.1016/j.jnutbio.2022.109257

Abstract: Cancer associated fibroblasts (CAFs) are one of the highly abundant components in the tumor microenvironment (TME). They secrete several cytokines, which amplified tumor progression, invasion, stemness, metastasis and angiogenesis. Here, we evaluate the potentiality of cytokines for the formation of cancer stem cells (CSCs) in oral cancer cells niche and investigate the anti-inflammatory and anti-carcinogenic effect of Resveratrol-nanoparticle (Res-NP). We first differentiated quiescent human fibroblasts into CAFs in vitro in response to PDGF-B and TGF-β stimulation and these CAFs were found to increase CXCL-12 and IL-6 secretion. CSCs enriched population was created by incubating H-357 cells with CAFs and cytokine-enriched CAFs-conditioned media (CAFs-CM). Likewise, CSCs-populated environment was also generated after incubating CAFs-CM to patient-derived primary oral cancer cells. It was noted that CXCL-12 and IL-6 secreted from CAFs significantly promoted CSCs growth, proliferation, aggressiveness, metastasis and angiogenesis. However, Res-NP reduced CSCs growth and proliferation by abrogating the secretion of CXCL-12 and IL-6. A significant decrease in the expression of metastatic and angiogenic markers, in ovo blood vascularization, intracellular NO generation, MMPs expression and tube formation was found upon Res-NP treatment. Reduction of representative CSCs and angiogenesis markers were also noted after Res-NP treatment in xenograft mice model. CXCL-12 physically interact with IL-6 and this interaction was diminished after Res-NP treatment. Moreover, the expression of CD133 and VEGF-A were downregulated either on Res-NP or CXCL-12/IL-6 specific inhibitors treated CSCs-enriched cells. Thus, the data suggest that CSCs growth is CXCL-12 and IL-6 dependent and Res-NP obstruct carcinogenesis and metastasis by inhibiting CXCL-12 and IL-6 production in in vitro, in vivo, in ovo and ex vivo systems.

2. Nayak, D., Paul, S., Das, C., Bhal, S., & Kundu, C. N. (2022). Quinacrine and Curcumin in combination decreased the breast cancer angiogenesis by modulating ABCG2 via VEGF A. Journal of Cell Communication and Signaling. https://doi.org/10.1007/s12079-022-00692-0

Abstract: Cancer stem cells (CSCs) cause drug resistance in cancer due to its extensive drug efflux, DNA repair and self-renewal capability. ATP binding cassette subfamily G member 2 (ABCG2) efflux pump afford protection to CSCs in tumors, shielding them from the adverse effects of chemotherapy. Although the role of ABCG2 in cancer progression, invasiveness, recurrence are known but its role in metastasis and angiogenesis are not clear. Here, using in vitro (CSCs enriched side population [SP] cells), ex vivo (patient derived primary cells), in ovo (fertilized egg embryo) and in vivo (patient derived primary tissue mediated xenograft (PDX)) system, we have systematically studied the role of ABCG2 in angiogenesis and the regulation of the process by Curcumin (Cur) and Quinacrine (QC). Cur + QC inhibited the proliferation, invasion, migration and expression of representative markers of metastasis and angiogenesis. Following hypoxia, ABCG2 enriched cells released angiogenic factor vascular endothelial growth factor A (VEGF A) and induced the angiogenesis via PI3K-Akt-eNOS cascade. Cur + QC inhibited the ABCG2 expression and thus reduced the angiogenesis. Interestingly, overexpression of ABCG2 in SP cells and incubation of purified ABCG2 protein in media induced the angiogenesis but knockdown of ABCG2 decreased the vascularization. In agreement with in vitro results, ex vivo data showed similar phenomena. An induction of vascularization was noticed in PDX mice but reduction of vascularization was also observed after treatment of Cur + QC. Thus, data suggested that in hypoxia, ABCG2 enhances the production of angiogenesis factor VEGF A which in turn induced angiogenesis and Cur + QC inhibited the process by inhibiting ABCG2 in breast cancer.

3. Girma, G., Panda, A.N., Roy, P.C., Ray, L., Mohanty, S & Chowdhary G. (2022). Molecular, biochemical, and comparative genome analysis of a rhizobacterial strain Klebsiella Sp. KBG6.2 imparting salt stress tolerance to Oryza sativa L. Environmental and Experimental Botany, 203, 105066. https://doi.org/10.1016/j.envexpbot.2022.105066

Abstract: One-third of global agricultural land is affected by salinity, thereby limiting the, productivity and sustainability of the cropping ecosystem. Rice is a globally important, economical crop but is largely affected by soil salinity, leading to the loss of standing crops. Plant growth-promoting rhizobacteria (PGPR) can be a potential solution to this problem by improving soil fertility, tolerance to stress and ultimately enhancing crop yield. However, the usage of PGPR in alleviating stress is limited and requires further investigation. In the current work, a PGPR strain, namely, Klebsiella sp. KBG6.2 was isolated from the rice rhizosphere of salt-affected agricultural fields in Odisha, India., The strain was found to be imparting salt tolerance to rice seedlings grown up to 200 mM of sodium chloride. The rice seedlings when treated with sodium chloride and coinoculated with Klebsiella sp. KBG6.2 showed better germination percentage, biomass, shoot, root length, chlorophyll content, and reducing sugar accumulation. The PGPR treatment also led to a reduction in reactive oxygen species (ROS) production, as evidenced by reduced ROS scavenging enzyme (catalase, superoxide dismutase, and peroxidase) activities and lower accumulation of the corresponding mRNA as, verified by real time PCR analysis. This was further supported by the reduced, accumulation of malondialdehyde which is an indicator of stress-induced oxidative damage. The genome sequence analysis of Klebsiella sp. KBG6.2 strain revealed, it to contain the required genes responsible for colonization and acclimation during stress conditions. The successful root colonization was further verified by scanning electron microscopy. Further, the obtained PGPR strain could provide saline stress tolerance to rice and serve as bioinoculant for sustainable agriculture in salt-affected areas.

4. Swalsingh, G., Pani, P., & Bal, N.C. (2022). Structural functionality of skeletal muscle mitochondria and its correlation with metabolic diseases. Clinical Science (Lond). 136(24):1851-1871. doi: 10.1042/CS20220636.

Abstract: The skeletal muscle is one of the largest organs in the mammalian body. Its remarkable ability to swiftly shift its substrate selection allows other organs like the brain to choose their preferred substrate first. Healthy skeletal muscle has a high level of metabolic flexibility, which is reduced in several metabolic diseases, including obesity and Type 2 diabetes (T2D). Skeletal muscle health is highly dependent on optimally functioning mitochondria that exist in a highly integrated network with the sarcoplasmic reticulum and sarcolemma. The three major mitochondrial processes: biogenesis, dynamics, and mitophagy, taken together, determine the quality of the mitochondrial network in the muscle. Since muscle health is primarily dependent on mitochondrial status, the mitochondrial processes are very tightly regulated in the skeletal muscle via transcription factors like peroxisome proliferator-activated receptor-γ coactivator-1α, peroxisome proliferator-activated receptors, estrogen-related receptors, nuclear respiratory factor, and Transcription factor A, mitochondrial. Physiological stimuli that enhance muscle energy expenditure, like cold and exercise, also promote a healthy mitochondrial phenotype and muscle health. In contrast, conditions like metabolic disorders, muscle dystrophies, and aging impair the mitochondrial phenotype, which is associated with poor muscle health. Further, exercise training is known to improve muscle health in aged individuals or during the early stages of metabolic disorders. This might suggest that conditions enhancing mitochondrial health can promote muscle health. Therefore, in this review, we take a critical overview of current knowledge about skeletal muscle mitochondria and the regulation of their quality. Also, we have discussed the molecular derailments that happen during various pathophysiological conditions and whether it is an effect or a cause.

5. Singh, T.D., Singh, N.I., Devi, K.M., Meiguilungpou, R., Khongsai, L., Singh, L.S., Bal, N.C., Swapana, N., Singh, C.B., & Singh, T.R. (2022). Planispine A Sensitized Cancer Cells to Cisplatin by Inhibiting the Fanconi Anemia Pathway. Molecules. 27(21):7288. doi: 10.3390/molecules27217288.

Abstract: The use of cisplatin as a chemotherapeutic drug is impeded by the development of drug resistance. Combination therapies of a chemosensitizer for cisplatin have been studied, but with little success, and the search for an effective combination therapy is continuing. Our earlier reports have shown that Zanthoxylum armatum DC. extract enhances the apoptotic effect of cisplatin in cancer cell lines. In this study, we purified and identified the bioactive phytocompound through bio-assay-guided purification, using column chromatography and HPLC. Chemical characterization using NMR and mass spectrometry revealed the compound as planispine A, with molecular structure C25H30O6 and molecular weight, 426.16 g/mol. Planispine A was found to inhibit cancer cell proliferation in a dose-dependent manner and to sensitize the cancer cells to cisplatin-augmented apoptotic cell death, in a caspase-dependent manner. A combination of planispine A and cisplatin induced S-phase cell cycle arrest, and reduced the expression of survival proteins such as cyclin D1. Interestingly, planispine A inhibits the Fanconi anemia pathway, as shown by reduced FANCD2 foci formation and FANCD2 monoubiquitination, which revealed the molecular mechanism of chemo-sensitization of cancer cells to cisplatin. Evaluation of this combination therapy in cisplatin-resistant tumors may lead to more efficient cisplatin treatment.

6. Pani, S., Dey, S., Pati, B., Senapati, U., & Bal, N.C. (2022). Brown to White Fat Transition Overlap With Skeletal Muscle During Development of Larger Mammals: Is it a Coincidence? Journal of Endocrinology Society. 6(12):bvac151. doi: 10.1210/jendso/bvac151.

Abstract: In mammals, adipose tissues and skeletal muscles (SkMs) play a major role in the regulation of energy homeostasis. Recent studies point to a possibility of dynamic interplay between these 2 sites during development that has pathophysiological implications. Among adipose depots, brown adipose tissue (BAT) is the major energy-utilizing organ with several metabolic features that resemble SkM. Both organs are highly vascularized, innervated, and rich in mitochondria and participate in defining the whole-body metabolic rate. Interestingly, in large mammals BAT depots undergo a striking reduction and concomitant expansion of white adipose tissue (WAT) during postnatal development that shares temporal and molecular overlap with SkM maturation. The correlation between BAT to WAT transition and muscle development is not quite apparent in rodents, the predominantly used animal model. Therefore, the major aim of this article is to highlight this process in mammals with larger body size. The developmental interplay between muscle and BAT is closely intertwined with sexual dimorphism that is greatly influenced by hormones. Recent studies have pointed out that sympathetic inputs also determine the relative recruitment of either of the sites; however, the role of gender in this process has not been studied. Intriguingly, higher BAT content during early postnatal and pubertal periods positively correlates with attainment of better musculature, a key determinant of good health. Further insight into this topic will help in detailing the developmental overlap between the 2 seemingly unrelated tissues (BAT and SkM) and design strategies to target these sites to counter metabolic syndromes.

7. Pani, P., & Bal, N.C. (2022). Avian adjustments to cold and non-shivering thermogenesis: whats, wheres and hows. Biological Reviews. 97(6):2106-2126. doi: 10.1111/brv.12885.

Abstract: Avian cold adaptation is hallmarked by innovative strategies of both heat conservation and thermogenesis. While minimizing heat loss can reduce the thermogenic demands of body temperature maintenance, it cannot eliminate the requirement for thermogenesis. Shivering and non-shivering thermogenesis (NST) are the two synergistic mechanisms contributing to endothermy. Birds are of particular interest in studies of NST as they lack brown adipose tissue (BAT), the major organ of NST in mammals. Critical analysis of the existing literature on avian strategies of cold adaptation suggests that skeletal muscle is the principal site of NST. Despite recent progress, isolating the mechanisms involved in avian muscle NST has been difficult as shivering and NST co-exist with its primary locomotory function. Herein, we re-evaluate various proposed molecular bases of avian skeletal muscle NST. Experimental evidence suggests that sarco(endo)plasmic reticulum Ca2+ -ATPase (SERCA) and ryanodine receptor 1 (RyR1) are key in avian muscle NST, through their mediation of futile Ca2+ cycling and thermogenesis. More recent studies have shown that SERCA regulation by sarcolipin (SLN) facilitates muscle NST in mammals; however, its role in birds is unclear. Ca2+ signalling in the muscle seems to be common to contraction, shivering and NST, but elucidating its roles will require more precise measurement of local Ca2+ levels inside avian myofibres. The endocrine control of avian muscle NST is still poorly defined. A better understanding of the mechanistic details of avian muscle NST will provide insights into the roles of these processes in regulatory thermogenesis, which could further inform our understanding of the evolution of endothermy among vertebrates.

8. Mandal, D., Patel, P., Verma, S.K., Sahu, B.R., Parija, T. (2022) Proximal discrepancy in intrinsic atomic interaction arrests G2/M phase by inhibiting Cyclin B1/CDK1 to infer molecular and cellular biocompatibility of d-limonene. Scientific Reports. 12(1):1-4. doi: https://doi.org/10.1038/s41598-022-21364-4

Abstract: The quest for different natural compounds for different biomedical applications especially in the treatment of cancer is at a high pace with increasing incidence of severity. D-limonene has been portrayed as one of the effective potential candidate centered to the context of breast cancer. The anticipation of its count as an effective biomedical agent required a detailed understanding of their molecular mechanism of biocompatibility. This study elucidates the mechanistic action of D-limonene channelized by the induction of apoptosis for controlling proliferation in breast cancer cells. The possible mechanism was explored through an experimental and computational approach to estimate cell proliferation inhibition, cell cycle phase distribution, apoptosis analysis using a flow cytometry, western blotting and molecular docking. The results showed reduced dose and time-dependent viability of MCF7 cells. The study suggested the arrest of the cell cycle at G2/M phase leading to apoptosis and other discrepancies of molecular activity mediated via significant alteration in protein expression pattern of anti-apoptotic proteins like Cyclin B1 and CDK1. Computational analysis showed firm interaction of D-limonene with Cyclin B1 and CDK1 proteins influencing their structural and functional integrity indicating the mediation of mechanism. This study concluded that D-limonene suppresses the proliferation of breast cancer cells by inducing G2/M phase arrest via deregulation of Cyclin B1/CDK1.

9. Mandal, D., Parija, T. (2022). Anticancer Mechanism of D-limonene: An Updated Review and Therapeutic Possibilities. Current Cancer Therapy Reviews. 2022;18(3):193-201. doi: https://doi.org/10.2174/1573394718666220421112750.

Abstract: Background: Despite remarkable advancement in screening, diagnosis, and treatment modalities cancer remains the second leading cause of death globally. Chemoprevention is considered to be a potential strategy for dealing with cancer incidence and mortality. The present demand for a less toxic well-tolerated natural anticancer compound that can be used for chemoprevention has drawn the attention towards D-limonene, which is a monocyclic monoterpene found in citrus oil. In recent years several in vitro and in vivo studies have supported the anticancer potential of D-limonene in various cancers. Although these studies have highlighted its course of action through immune modulation, anti-oxidant activity, apoptosis, autophagy, etc. various scientific pieces of evidence support the fact that it targets multiple pathways to inhibit cancer. Objective: The current review focuses on the molecular mechanisms underlying the anticancer activity of D-limonene and discusses its potential to be used as a cost-effective chemopreventive and chemotherapeutic drug alone or in combination with other drugs. Methods Scientific databases like Web of Science, Scopus, PubMed, Google Scholar, PubMed Central etc have been used to review new and recent insights into the anticancer mechanism of D-limonene. Conclusion: In this review, we discussed the overall, significance of the anticancer mechanisms of D-limonene which include modulation of apoptosis, promotion of autophagy and inhibition of angiogenesis and metastasis. It also inhibits oncogenic signaling molecules and related transcription factors. Additionally, it also acts in combination with other anticancer compounds to inhibit cancer.

10. Panda, C., & Mahapatra, R. K. (2022). An update on cerebral malaria for therapeutic intervention. Molecular biology reports, 49(11), 10579–10591. doi: 10.1007/s11033-022-07625-5

Abstract: Background: Cerebral malaria is often pronounced as a major life-threatening neurological complication of Plasmodium falciparum infection. The complex pathogenic landscape of the parasite and the associated neurological complications are still not elucidated properly. The growing concerns of drug resistant parasite strains along with the failure of anti-malarial drugs to subdue post-recovery neuro-cognitive dysfunctions in cerebral malaria patients have called for a demand to explore novel biomarkers and therapeutic avenues. Due course of the brain infection journey of the parasite, events such as sequestration of infected RBCs, cytoadherence, inflammation, endothelial activation, and blood-brain barrier disruption are considered critical. Methods: In this review, we briefly summarize the diverse pathogenesis of the brain-invading parasite associated with loss of the blood-brain barrier integrity. In addition, we also discuss proteomics, transcriptomics, and bioinformatics strategies to identify an array of new biomarkers and drug candidates. Conclusion: A proper understanding of the parasite biology and mechanism of barrier disruption coupled with emerging state-of-art therapeutic approaches could be helpful to tackle cerebral malaria.

11. Sinha, A., Simnani, F. Z., Singh, D., Nandi, A., Choudhury, A., Patel, P., Jha, E., Chouhan, R. S., Kaushik, N. K., Mishra, Y. K., Panda, P. K., Suar, M., & Verma, S. K. (2022). The translational paradigm of nanobiomaterials: Biological chemistry to modern applications. Materials today. Bio, 17, 100463. doi: 10.1016/j.mtbio.2022.100463

Abstract: Recently nanotechnology has evolved as one of the most revolutionary technologies in the world. It has now become a multi-trillion-dollar business that covers the production of physical, chemical, and biological systems at scales ranging from atomic and molecular levels to a wide range of industrial applications, such as electronics, medicine, and cosmetics. Nanobiomaterials synthesis are promising approaches produced from various biological elements be it plants, bacteria, peptides, nucleic acids, etc. Owing to the better biocompatibility and biological approach of synthesis, they have gained immense attention in the biomedical field. Moreover, due to their scaled-down sized property, nanobiomaterials exhibit remarkable features which make them the potential candidate for different domains of tissue engineering, materials science, pharmacology, biosensors, etc. Miscellaneous characterization techniques have been utilized for the characterization of nanobiomaterials. Currently, the commercial transition of nanotechnology from the research level to the industrial level in the form of nano-scaffolds, implants, and biosensors is stimulating the whole biomedical field starting from bio-mimetic nacres to 3D printing, multiple nanofibers like silk fibers functionalizing as drug delivery systems and in cancer therapy. The contribution of single quantum dot nanoparticles in biological tagging typically in the discipline of genomics and proteomics is noteworthy. This review focuses on the diverse emerging applications of Nanobiomaterials and their mechanistic advancements owing to their physiochemical properties leading to the growth of industries on different biomedical measures. Alongside the implementation of such nanobiomaterials in several drug and gene delivery approaches, optical coding, photodynamic cancer therapy, and vapor sensing have been elaborately discussed in this review. Different parameters based on current challenges and future perspectives are also discussed here.

12. Bhattacharjee, R., Nandi, A., Sinha, A., Kumar, H., Mitra, D., Mojumdar, A., Patel, P., Jha, E., Mishra, S., Rout, P. K., Panda, P. K., Suar, M., & Verma, S. K. (2022). Phage-tail-like bacteriocins as a biomedical platform to counter anti-microbial resistant pathogens. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 155, 113720.DOI: 10.1016/j.biopha.2022.113720

Abstract:Phage Tail Like bacteriocins (PTLBs) has been an area of interest in the last couple of years owing to their varied application against multi-drug resistant (MDR), anti-microbial resistant (AMR) pathogens and their evolutionary link with the dsDNA virus and bacteriophages. PTLBs are defective phages derived from Myoviridae and Siphoviridae phages, PTLBs are distinguished into R-type (Rigid type) characterized by a non-flexible contractile nanotube resembling Myoviridae phage contractile tails, and F-type (Flexible type) with a flexible non-contractile rod-like structure similar to Siphoviridae phages. In this review, we have discussed the structural association, mechanism, and characterization of PTLBs. Moreover, we have elucidated the symbiotic biological function and application of PTLBs against MDR and XDR pathogens and highlighted the evolutionary role of PTLBs. The difficulties that must be overcome to implement PTLBs clinically are also discussed. It is imperative that these issues be addressed by academics in future studies before being implemented in clinical settings. This article is novel in its way as it will not only provide us with a gateway that acts as a novel strategy for scholars to mitigate and control the uprising issue of AMR pathogens but also promote the development of clinical studies for PTLBs.

13. Behera, H. T., Mojumdar, A., Behera, S. S., Das, S., & Ray, L. (2022). Biocontrol of wilt disease of rice seedlings incited by Fusarium oxysporum through soil application of Streptomyces chilikensis RC1830. Letters in applied microbiology, 75(5), 1366–1382. doi: 10.1111/lam.13807

Abstract:The genus Streptomyces includes many antifungal metabolite-producing novel strains. Fusarium oxysporum soil-inhabiting pathogenic fungi, that affects rice to cause wilt disease. This work demonstrates the efficacy of novel Streptomyces chilikensis strain RC1830, previously isolated from estuarine habitat Chilika Lake in preventing the F. oxysporum wilting/root rot disease and promoting the growth of rice (Var. Swarna) seedlings. A total of 25 different compounds were identified from crude extracts of S. chilikensis RC1830 by GC-MS. In pot trial experiments, Streptomyces-treated rice seedlings showed significantly reduced disease severity index by 80·51%. The seedlings growth parameters (root length, root fresh weight and root dry weight) were also increased by 53·91, 62·5 and 73·46%, respectively, in Streptomyces-treated groups of seedlings compared to Fusarium-infected seedlings. Similarly, the shoot length, shoot dry weight and shoot fresh weight were also increased by 26, 58 and 34·4%, respectively, in Streptomyces-treated groups of seedlings compared to Fusarium-infected seedlings. Formulations of the strain were prepared using seven organic and inorganic wastes as the carrier material and the shelf lives of the propagules were also monitored. Vermiculite and activated charcoal formulations stored at 4°C exhibited a higher viable cell count after 3 months of storage.

14. Pattanaik, K. P., Sengupta, S., Jit, B. P., Kotak, R., & Sonawane, A. (2022). Host-mycobacteria conflict: Immune responses of the host vs. the mycobacteria TLR2 and TLR4 ligands and concomitant host-directed therapy. Microbiological research, 264, 127153. doi: 10.1016/j.micres.2022.127153

Abstract:Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is one of the most grievous infectious diseases with long-term morbidity and unpredicted mortality rates globally. Thus, understanding the host-pathogen interactions to develop potential drugs is the most focused area of research. Mtb has many antigens communicating with host cells via various pattern recognition receptors (PRRs). From which, toll-like receptors-2 and 4 (TLR2 and 4) are two major PRRs that provide the primary immune response to Mtb infection of the respiratory tract. As a result, the TLR-mycobacterium antigen interaction triggers a variety of crucial innate immune signalling mechanisms such as phagosome maturation, oxidative stress, elicitation of cell deaths, production of proinflammatory cytokines, and eventually associates with the adaptive immune response to establish infection. Despite the extensive investigations on TLR2 and 4 Mtb ligands that have a significant role in the immune defence system, there are still many unsolved concerns driving researchers to explore the obscures. This review focuses on the host immune modulation due to Mtb-TLR2 and 4 ligand interaction. Subsequently, the host TLR2 and 4 immune signals in cooperation with other PRRs and successive cytokine expressions are discussed. Also highlighted are some recent findings on host-directed therapy related to TLRs that aid in developing novel immunotherapeutic prospects for the better control of Mtb infection.

15. Mohanty, R., Manoswini, M., Dhal, A. K., & Ganguly, N. (2022). In silico analysis of a novel protein in CAR T- cell therapy for the treatment of hematologic cancer through molecular modelling, docking, and dynamics approach. Computers in biology and medicine, 151(Pt A), 106285. odi: 10.1016/j.compbiomed.2022.106285

Abstract: Cellular therapy has emerged as a key tool in the treatment of hematological malignancies. An advanced cell therapy known as chimeric antigen receptor T cell therapy (CAR T-cell therapy) has been approved by the United States Food and Drug Administration (FDA) as KYMRIAH by Novartis and YESCARTA by Gilead/Kite pharma in the year 2017. A chimeric receptor is composed of an extracellular antigen recognition site along with some co-stimulating and signalling domains. On the whole, it turns out to be one of the most potent receptors on T cells targeting a specific type of cancer cell based on its antigenic marker. CD19 CAR T-cell therapy is the first clinically approved therapy for lymphoma with remarkable results in complete remission of B cell lymphoblastic leukemia up to 90%. The high rate of effectiveness of the CAR T-cell therapy against B-ALL justifies the investigation and application of this therapy for fatal diseases like all types of hematological malignancies. The most critical aspect of chimeric receptor therapy is designing and building an artificial receptor that is specific to a given type of cancer. For this reason, the in silico technique is an appropriate model to investigate the integrity and effectiveness of the engineered chimeric receptor prior to commencing in vitro experiments followed by clinical trials. This computerized experimental study aids in predicting the molecular mechanism of chimeric protein and how it interacts with both ligands. We have anticipated various features of the chimeric protein in terms of qualitative analysis (structure, protein modelling, physiological properties) and functional analysis (antigenicity, allergenicity, its receptor-ligand binding ability, involving signalling pathways). Furthermore, the reliability and validation of the binding mode of the chimeric protein against receptors were performed through a complex molecular dynamics simulation for a 100 ns timeframe in an aqueous environment. The obtained simulation study showed that CD30 was a better approachable marker as compared to CD20 due to its better binding energy score and also binding conformations stability.

CONFERENCE ARTICLES

1. Thakur, C.; Karthikeyan, C.; Bhal, S.; Kundu, C.; Moorthy, N. (2022). Targeted Delivery of Doxorubicin to Breast Cancer Cells by Multiwalled Carbon Nanotubes Functionalized with Lysine via 1,3-Dipolar Cycloaddition and Conjugation with Sugar moieties. Proceedings of the 8th International Electronic Conference on Medicinal Chemistry, 1–30 November 2022, https://doi.org/10.3390/ECMC2022-13421

Abstract: Multiwalled carbon nanotubes (MWCNTs) have gained a lot of multidisciplinary attention and biomedical applications such as drug delivery, because of their distinct physicochemical characteristics. Even though MWCNTs are not used because of low dispersibility in aqueous or non-aqueous medium. Functionalizing MWCNTs is an attractive way to overcome this drawback, it improves biocompatibility and promotes ligand attachment for targeted drug delivery. However, most functionalization techniques include hazardous procedures and costly chemicals. The current study uses a straightforward, economically advantageous method to functionalize MWCNTs with lysine through 1,3-dipolar cycloaddition for enhanced dispersibility and to offer a ligand anchoring ε-amino group for targeted delivery to breast cancer. MWCNTs had been functionalized with lysine and sugar moieties ligands (galactose/mannose) to create efficient nanocarriers that can bind to lectin receptors in MDA-MB-231 or MCF-7 cancer cells. Doxorubicin (Dox) was loaded into the ligands conjugated MWCNTs. In comparison to pristine MWCNTs, 1,3-lysinated MWCNTs conjugated with ligands demonstrated enhanced dispersion in an aqueous medium and greater drug loading capacity. Drug release studies in pH 7.4 were 20% only & in pH 5.0 was around 75%. Dox-loaded MWCNTs provided pH-dependent releases and enhanced Dox accumulation inside the cancer cells, as evidenced by higher inhibition of MDA-MB-231 or MCF-7 compared to plain Dox, and unloaded Dox MWCNTs nanoformulations provided negligible cytotoxicity. Based on the results, MWCNTs functionalized with lysine by 1,3-dipolar cycloaddition offer promising nontoxic nanoplatforms with improved aqueous dispersibility and potential for conjugation with ligands for targeted delivery of Dox to breast cancer cells.

2. Singh, S.; Paul, S.; Karthikeyan, C.; Brás, N.; Kundu, C.; Moorthy, N. (2022). Design, synthesis and biological evaluation of Novel 1H-benzo[d]imidazole derivatives as Fatty Acid Synthase (FASN) inhibitors for cancer treatment. Proceedings of the 8th International Electronic Conference on Medicinal Chemistry, 1–30 November 2022. https://doi.org/10.3390/ECMC2022-13414

Abstract: FASN a metabolic oncoprotein overexpressed in multiple cancer and regulates the fatty acid requirement for proliferated cells. Thus, FASN has been proposed as a promising novel target for anticancer drug discovery. Herein we report the de-novo design and synthesis of small molecule FASN inhibitors (CTL) targeting breast and colorectal cancer. The structure-activity relationship studies led to identify CTL-1 and CTL-7 as potent, selective FASN inhibitors had an IC50 2.5 and 3.0 µM. The CTL-1 and CTL-7 inhibits proliferation of colon cell (with IC50 range of 3-5 µM) in HCT-116, CaCO2 and breast cells MCF-7, MDA-MB-231. However, in non-cancerous cell line HEK-293 the IC50 of CTL-1 and CTL-7 was above 30 µM. Further cell cycle analysis and apoptosis assay of CTL-1 and CTL-7 in HCT-116 cells represents S-phase arrest along with prolong apoptotic effect. The western blot analysis of CTL-1 and CTL-7 establish the FASN pathway participation in causing cell apoptosis. The molecular dynamics simulation studies indicate the high affinity of CTL-1 and CTL-7 against the FASN enzyme.

PATENT FILED/PUBLISHED/GRANTED

1. Dr. Chinmayee Sethy, Dr. Chanakya Nath Kundu. Patent title: Nectin-4 as a Soluble Biomarker for The Detection of Cancer (India Patent Application no. 202231059837)

Abstract: The present invention relates to methods for detecting and diagnosing cancer Moreover, the present invention relates to methods for screening an agent for or preventing cancer. More Particularly the present invention relates to a development of biomarker for the detection of various cancers. The very well-expressed protein (namely Nectin-4) in all types of cancers and this protein specifically expressed in cancer cells but not in any normal tissue, cells in the body. Moreover, the aggressiveness of cancer can be predicted or determined by checking the level of Nectin-4 expression in cancer samples. The Nectin-4 expression can be checked in two easily available samples such as blood and lymph node aspirates. No robust and complex techniques are involved in the sample collection, processing, and detection of the level of Nectin-4 expression. Results showed that the level of Nectin-4 expression can help in deciding the treatment plan for effective treatment.

2. Ms. Mamata Das, Mr. Biswajit Das, Dr. Chanakya Nath Kundu. Patent title: Polysaccharide-Based Wound Healing Patch. (India Patent Application no. 202231059838)

Abstract: The present invention relates to composition of a curcumin-loaded carboxymethyl tamarind (CMT) /crystalline nanocellulose (CNC) /nano silver-based wound healing patch to promote the wound healing process more rapidly with potential antibacterial activity. The developed patch contains naturally available ingredients to avoid cytotoxicity of the patch. The developed patch is mechanically flexible with excellent antibacterial activity and wound healing potential. Formulated patch can avoid the unusual excess dressing changes during the healing process as compared to conventional dressing materials.

3. Dr. Shakti Ranjan Satapathy, Mr. Somya Ranjan Dash, Dr. Chanakya Nath Kundu. Patent title: Quinacrine-gold hybrid nanoparticle and NIR irradiation for treatment of oral cancer. (India Patent Application no. 202231059841)

Abstract: The present invention relates to preparation of hybrid nanoparticles. we use a natural bioactive compound, Quinacrine (QC) which specifically target and kill the cancer cells. Further, QC has been Quinacrine is conjugated with a metallic gold nanoparticle (AuNP). we mix both of them to generate a hybrid nanoparticle (QAuNP) which when excited with a fixed laser power of NIR offers excellent anticancer and photothermal potentiality at the localized oral cancer by generating heat. The modest heat produced by hybrid nanoparticles activated with NIR can destroy oral cancer cells while protecting adjacent healthy cells from heat stress. The developed hybrid nanoparticle poses no threat to the body’s healthy cells. Due to its reduced size of, it demonstrated an improved permeability and retention effect (EPR effect).

4. Applicants: Gopal Chowdhary, Berhanu Girma and Pamela Chanda Roy

Title of the invention: A non-genetically modified approach of use of rhizobacterial cocktail for imparting tolerance for abiotic stress matrix in crop plants

Indian Patent application number- 202231060862

Web link: https://ipindiaservices.gov.in/PublicSearch/PublicationSearch/ApplicationStatus

Abstract: Abiotic stress is a serious threat to crop plants. Due to ongoing climate change, it has become imperative to find ways to counter this. Here we have devised a mechanism to use a cocktail of plant growth-promoting rhizobacteria to alleviate the abiotic stress in crop plants.

Applicants: Dr. D. Mandal, Dr. B. Panigrahi and Dr. R. Singh.

Title of the invention: Metal based Hybrid Protease Inhibitor for Multiple Proteases

India patent application no. 202131010934, patent published.

Abstract: The present invention describes a metal-based hybrid multi-protease inhibitor that uses the organic-inorganic hybrid combination of flavonoids and metal as an inhibitor that aids to inhibit multiple proteases and has a wide range of inhibition efficiency. The proposed hybrid protease inhibitor provides significant inhibition against a broad range of proteases such as trypsin, chymotrypsin, proteinase K, and pepsin. The protease inhibitor is compatible and active in different buffer conditions. The hybrid protease inhibitor is non-toxic, water-soluble, and non-hazardous to the patient. Further, the proposed hybrid inhibitor is a low-cost hybrid protease inhibitor that is 10 times cheaper than commercially available inhibitors. The hybrid protease inhibitor has various applications across a wide range of industries including as an anti-aging cream and an antidandruff shampoo in the cosmetic industry, as a pest control agent in agriculture, as an antiviral drug in the pharmaceutical industry, protein-drug purification industry, and biotech industry.

RESEARCH FUNDING

Grant or Project Awarded (Both Sponsored Research and Consultancy):

Title of the Sanctioned Project: Human host candidate genetic marker revelation by transcriptomic profiling of vitamin D deficient Plasmodium vivax and Plasmodium falciparum malaria cases in the paediatric population of Odisha, India

Principal Investigator’s Name: Dr. Biswadeep Das, School of Biotechnology

Co-Principal Investigator’s Name: Prof. Sanghamitra Pati, Dr. Chinmay Behera, Dr. RajaniKanta Mahapatra

Name of Funding Agency: ICMR-MERA, Govt. of India

Sanctioned Amount of Project: Rs. 56 Lakhs for 2 years

Abstract of the Project:

Malaria is the most important vector borne disease in various parts of India, in particular regions with hot and humid climates, and is caused by the parasite, Plasmodium species. Because vitamin D deficient malaria cases mostly occur in most of the children in the age group of 2-17 years, it is important to understand the underlying genetic mechanisms associated with vitamin D deficiency and vivax malaria. This research will involve transcriptomic profiling of vitamin D deficient malaria cases in the pediatric population of Odisha, India for the determination of specific host genetic markers. The data generated will assist in elucidating the role of vitamin D in malaria outcome, that could assess the genetic basis of malaria severity in vitamin D deficient cases, which could be utilized by malariologists to take a call to provide vitamin D supplementation for ameliorating malaria symptoms.

Ph.D. DEGREE AWARDED (PROVISIONAL)

Student’s Name: Dr. Deepika Nayak

Supervisor’s Name: Dr. Chanakya Nath Kundu, School of Biotechnology

Thesis Title: Study the role of Quinacrine and Curcumin combination for anti-breast cancer progression by deregulation of ABCG2 in pre-clinical model systems

Abstract of the Thesis:

Cancer stem cell like cells (CSCs) present a challenge in the management of cancers due to their involvement in the development of resistance against various chemotherapeutic agents. Over expression of ABCG2 transporter gene is one of the factors responsible for drug resistance in CSCs, which causes efflux of therapeutic drugs from these cells. Cancer stem cells (CSCs) cause drug resistance in cancer due to its extensive drug efflux, DNA repair and self-renewal capability. ATP binding cassette subfamily G member 2 (ABCG2) efflux pump afford protection to CSCs in tumors, shielding them from the adverse effects of chemotherapy. The development of inhibitors against CSCs has not achieved any significant success, till date. In this work, we have evaluated the anti-proliferative activity of curcumin (Cur) and quinacrine (QC) against CSCs using in vitro model system. Cur and QC synergistically inhibited the proliferation, migration and invasion of CSCs enriched side population (SP) cells of cigarette smoke condensate induced breast epithelial transformed (MCF-10A-Tr) generated metastatic cells. Cur+QC combination increased the DNA damage and inhibited the DNA repair pathways in SP cells. Uptake of QC increased in Cur pre-treated SP cells and this combination inhibited the ABCG2 activity by the reduction of ATP hydrolysis in cells. In vitro DNA binding reconstitution system suggests that QC specifically binds to DNA and caused DNA damage inside the cell. Decreased level of ABCG2, representative cell survival and DNA repair proteins were noted after Cur+QC treatment in SP cells. The molecular docking studies were performed to examine the binding behaviour of these drugs with ABCG2, which showed that QC (-53.99 kcal/mol) and Cur (-45.90 kcal/mol) occupy a highly overlapping interaction domain. This suggested that in Cur pre-treated cells, the Cur occupied the ligand-binding site in ABCG2, thus making the ligand binding site unavailable for the QC. This causes an increase in the intracellular concentration of QC. The results indicate that Cur+QC combination causes CSCs death by increasing the concentration of QC in the cells and thus causing the DNA damage and inhibiting the DNA repair pathways through modulating the ABCG2 activity. Although the role of ABCG2 in cancer progression, invasiveness, recurrence are known but its role in metastasis and angiogenesis are not clear. Here, using in vitro (CSCs enriched side population [SP] cells), ex vivo (patient derived primary cells), in ovo (fertilized egg embryo) and ex vivo (patient derived primary tissue mediated xenograft (PDX)) system, we have systematically studied the role of ABCG2 in angiogenesis and the regulation of the process by Curcumin (Cur) and Quinacrine (QC). Cur+QC inhibited the proliferation, invasion, migration and expression of representative markers of metastasis and angiogenesis. Following hypoxia, ABCG2 enriched cells released angiogenic factor vascular endothelial growth factor A (VEGF A) and induced the angiogenesis via PI3K-Akt-eNOS cascade. Cur+QC inhibited the ABCG2 expression and thus reduced the angiogenesis. Interestingly, overexpression of ABCG2 in SP cells and incubation of purified ABCG2 protein in media induced the angiogenesis but knockdown of ABCG2 decreased the vascularization. In agreement with in vitro results, ex vivo data showed similar phenomena. An induction of vascularization was noticed in PDX mice but reduction of vascularization was also observed after treatment of Cur+QC. Thus, data suggested that in hypoxia, ABCG2 enhances the production of angiogenesis factor VEGF A which in turn induced angiogenesis and Cur+QC inhibited the process by inhibiting ABCG2 in breast cancer.

2. Student’s Name: Dr. Berhanu Girma

Supervisor’s Name: Dr. Gopal Chowdhary, School of Biotechnology

Thesis Title: Isolation and Characterization of Plant Growth-Promoting Rhizobacteria from Rice Fields of Konark and Investigating their Efficacy in Imparting Abiotic Stress Tolerance

Abstract of the Thesis:

Abiotic stress is a major bottleneck in agricultural productivity. It is estimated that 20% world’s cultivated land and 33% of the total irrigated agricultural land are salt-affected and, it is increasing at an annual rate of 10% and as per an estimate, about 50% of agricultural land will be under the impact of saline stress by 2050 (Tanji, 1990; Jamil et al., 2011).In the Indian scenario, about 7 million hectares of agricultural land are affected by saline stress, which primarily lies in the states of West Bengal, Odisha, Andhra Pradesh, Pondicherry, Tamil Nadu, Kerala, Karnataka, Maharashtra, Gujarat, and Goa (Abrol and Bhumbla, 1973). This phenomenon is further aggravated by ongoing global climate change. Maintaining crop productivity under salt stress would require additional efforts (Turral et al., 2011). Soil salinization and the unanticipated threat of climate change scenarios are two of modern agriculture’s primary concerns. Due to a decrease in the yield because of saline stress conditions, the application of an excessive amount of chemical fertilizer has become a common norm, which in turn leads to environmental degradation. This has led to the search for alternate options for imparting saline stress tolerance to agriculturally important crops and maintaining the yield. Recent research has demonstrated the use of plant growth-promoting rhizobacteria (PGPR) can improve the tolerance of crop plants against abiotic stresses (Gupta et. al., 2000; Jha et al., 2012; Rajput et al., 2013; Sapre et al., 2018; Sijilmassi et al., 2020). The investigation of the PGPR strain’s whole-genome data, as well as the identification of their plant growth-promoting and stress-tolerance genes, is critical since it will enhance our understanding of molecular mechanisms in the development of better bio-fertilizers using PGPR-assisted technology (Schuster, 2008). As explained above, the state of Odisha has a long coastline and saline stress is a major concern for agriculture. Further, rice is a major crop being cultivated in the state of Odisha. Hence, we intended to isolate the PGPR strain/s which could impart saline stress tolerance to rice plants. For this, we have used the rhizospheric soil from the Konark area of the state of Odisha (19°53ʹ14.80ʺN, 86°05ʹ40.55ʺE). The area under investigation lies in the vicinity of the sea and hence has a persistent problem of saline stress. No previous studies on the PGPR from this area have been reported. The following objectives for the research work were formulated:
Isolation and characterization of PGPR strain/s from the salt-affected rhizospheric area of paddy fields (Konark, Odisha, India)
To analyze the whole genome sequence of PGPR strain/s and present a comparative genome snapshot to understand the PGPR functional genes that promote plant growth and impart saline stress tolerance to plants
To investigate the efficacy of isolated PGPR strain/s in imparting the saline stress tolerance to rice seedlings (Oryza sativa L.).

3. Student’s Name: Dr. Swatishree Sradhanjali

Supervisor’s Name: Dr. Mamatha M. Reddy, LVPEI (Supervisor) and Dr. Rahul Modak, KIIT Deemed to be University (Co-supervisor)

Thesis Title: The role of MYCN in regulating metabolic reprogramming in retinoblastoma.

Abstract of the Thesis:

MYCN is a master regulator of several cellular processes including metabolism. It is found to be overexpressed and is linked to dysregulated metabolism in several cancers. Therefore, a thorough understanding of metabolic processes and the targeting of metabolic enzymes to control particular bioenergetic activities in cancer cells is of paramount importance. Cancer cells rely on increased bioenergetics for rapid cell growth and proliferation. Several scientific studies have suggested that metabolic reprogramming is a key to cancer aggressiveness possibly via activating numerous growth signalling pathways. Further, MYC overexpression/amplification has also been linked to aggressive phenotype and increased proliferation rates in several malignancies. Currently, the mechanism behind metabolic reprogramming in adult cancers has been studied widely. However, altered metabolism in pediatric cancers such as retinoblastoma (RB) is just beginning to emerge. The predictive importance of mitochondrial OXPHOS, association between lipid metabolism and invasion has been reported in RB. However, the mechanism behind MYCN regulation of metabolic pathways in RB was yet to be deciphered until this thesis. Consequently, in the current thesis, our findings show that MYCN is overexpressed in retinoblastoma (RB) and regulates various metabolic pathways. We sought to decipher the significance of two important metabolic genes regulated by MYCN namely pyruvate dehydrogenase kinase (PDK)1 and 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB) 3 in RB and targeted these enzymes using small molecule inhibitors or by shRNA approach. Interestingly, we found that targeted inhibition of PDK1 and PFKFB3 resulted in decreased cell viability and migration of RB cells compared to the untreated controls. Further, it also led to increased apoptosis and cell cycle arrest. The effects of PDK1 and PFKFB3 inhibition on metabolic parameters such as glucose uptake and lactate levels were also studied and found to be decreased. Overall, the above results suggest that these genes could be potential therapeutic targets in RB. Additionally, we also tested the therapeutic efficacy of first-line chemotherapeutic drugs such as melphalan and carboplatin in combination with small molecule inhibitors of MYCN, PDK1 and PFKFB3 on RB cell lines and synergistic drug interactions were observed. However, some levels of additive and antagonist interaction between the drugs were also seen. These drug combination study provided a rationale for developing additional targeted therapeutic strategies for RB. However, comprehensive in vivo studies are required to understand and elucidate the synergistic growth inhibitory effects of these drugs in combination. The findings presented in this thesis show that targeting MYCN and MYCN-regulated metabolic pathways may be an interesting basis for developing innovative treatment approaches for retinoblastoma. Further, targeted inhibition of MYCN and metabolism may possibly improve the therapeutic efficacy of the chemotherapeutic agents in combination in MYCN driven retinoblastomas.

4. Student’s Name: Dr. Sourav Mishra

Supervisor’s Name: Dr. Dindyal Mandal, School of Biotechnology

Thesis Title: Exploring Catechol in nanoparticle synthesis and potential applications of catechol generated nanoparticles.

Abstract of the Thesis:

Catechol is demonstrated as a versatile reducing agent/stabilizing agent for the facile and one pot synthesis of monometallic (Au, Se and Pd) and bimetallic nanoparticles in alkaline condition (pH = 11) based on the redox chemistry of catechol. Catechol-derived PdNPs and SeNPs exhibit laccase and peroxidase like activity, respectively. Catechol derived gold nanoparticles serving as an excellent support for enzyme immobilization with recyclable properties are also reported here. As a test model protienase k immobilized on gold nanoparticles exhibits significant biocatalytic activities without compromising the enzyme activity. In summary. this work offers a versatile reducing agent for nanoparticle synthesis and demonstrates new generation of artificial enzymes. Herein, a label-free, fast, colorimetric dual sensing system based on the surface chemistry of catechol-generated gold nanoparticles (cAuNPs) is reported with very low limit of detection. In this study, a facile catechol mediated gold nanoparticle (cAuNPs) synthesis has been established and characterised by UV-vis spectroscopy, dynamic light scattering (DLS) techniques,high resolution transmission electron microscopy (HR-TEM) and Fourier transform infrared spectroscopy (FTIR). The newly generated cAuNPs exhibit plasmonic response toward the industrially important rare earth element ytterbium (Yb³⁺) and biologically significant enzyme proteinase k (PK). The strong affinity of the AuNPs toward the target analyte (Yb³⁺, PK) is associated with the rapid colorimetric changes, distinct spectral shift and formation of larger size of gold nanoparticles. The results highlight the potential application of cAuNPs as rapid, cost-effective and sensitive colorimetric sensor for Yb³⁺ and proteinase k (PK). Catechol generated palladium nanoparticles (PdNPs) have been utilized for the selective degradation of anionic dyes in aqueous solution. Palladium nanocatalyst (4μg/ml) exhibits complete degradation of anionic azo dye congo red (CR) (150 μM) within 10 minutes of reaction time. The degradation efficiency is found to be linearly dependent on the initial dye concentration, PdNP catalyst concentration, temperature and pH of the solution. The degradation efficiency increases with the decrease in pH of the solution and increasing PdNP catalyst concentration. Furthermore, with respect to the dye concentration, the degradation rate is found to be first order reaction kinetics. The recyclability of the catalyst confirms that even after 3^rd cycle 50% efficiency of the catalyst has been well retained and the degradation efficiency is found to be 92%. Gas chromatography-mass spectrometry (GC-MS) analysis of the degraded products such as benzidine reveals the degradation through reductive cleavage of azo linkage. In summary, the results highlight the potential development of a nanocatalyst for the selective removal of anionic azo dye from the wastewater.

NATIONAL/INTERNATIONAL AWARD RECEIVED BY THE FACULTY MEMBER:

Name of the Faculty: Dr. Chanakya Nath Kundu

Title of the Award: Plenary Session Award

Award Agency: Indian Association of Biomedical Scientists (IABMS) 43rd Annual Conference of Indian Association of Biomedical Scientists & International Conference on “An Integrative Approach Towards Health Protection and Health Promotion” in December, 2022.

Abstract:

Cancer stem cells secrete diffusible factors into the microenvironment that bind to specific endothelial cell receptors and initiate the angiogenesis, metastasis and cancer growth. Tumor- induced angiogenesis and metastasis are important parameter of tumorigenesis and is critical for tumor growth and metastasis. A pvrl-4 encoded gene, NECTIN-4, has potential roles in cancer cell growth and aggressiveness, and it is only expressed in cancer cells. There is evidence that nectin-4 plays a role in tumorigenesis, but the function of nectin-4 in tumor metastasis and angiogenesis has lacked thorough evidence of mechanism. It comprise of three specific domain endo ( N-terminal), ecto ( C-terminal) and trans membrane domain. The N-terminal domain of the nectin-4 induced the DNA repair and C-terminal domain is responsible for angiogenesis. Using highly metastatic cancer cells and human umbilical vein endothelial cells (HUVECs), we have developed an excellent angiogenesis model and systematically studied the contribution of nectin-4 to angiogenesis. We also provide in-depth in vitro, in ovo, ex vivo and in vivo evidence that nectin-4 causes angiogenesis and propose that nectin-4 is an angiogenesis biomarker in breast cancer. Following hypoxia, the expression of ADAM-17 in metastatic breast cancer stem cells (mBCSCs) causes the shedding of the ecto-domain of nectin-4 into the microenvironment, which physically interacts with integrin-β4 specifically on endothelial cells. This interaction promotes angiogenesis via the Src, PI3K, AKT, iNOS pathway and not by Phospho-Erk or NF- κβ pathways. In vitro, in ovo and in vivo induction and abrogation of an angiogenesis cascade in the presence and absence of the nectin-4 ecto-domain, respectively, confirms its role in angiogenesis. Thus, disrupting the interaction between nectin-4 ecto-domain and integrin-β4 may provide a means of targeting mBCSC-induced angiogenesis.

8. ANY NATIONAL/INTERNATIONAL AWARD OR PUBLICATION (R&D RELATED) RECEIVED BY STUDENTS:

Name of the Student: Mr. Saptarshi Sinha

Title of the Award: Best Paper Presenter Award (Oral Presentation Category)

Award Agency: Indian Association of Biomedical Scientists (IABMS) at 43rd Annual Conference of Indian Association of Biomedical Scientists & International Conference on “An Integrative Approach Towards Health Protection and Health Promotion” in December, 2022.

Abstract:

Background Although sensitization of BRCA-mutated, homologous recombination (HR)-deficient breast cancer cells through PARP inhibitor is widely studied, not much is known about the treatment of BRCA-wild-type, HR-proficient breast cancer.

Objectives Our aim is to investigate whether a natural bioactive compound, Resveratrol (RES), can induce DNA double-strand breaks in HR-proficient breast cancer cells and Olaparib (OLA), a PARP inhibitor, can enhance the RES-mediated apoptosis by deregulating the HR repair pathway.

Methods The detailed mechanism of anti-cancer action of RES+OLA combination in breast cancer has been evaluated using in vitro (MCF-7 and T47D cell lines), ex vivo (patient-derived primary breast cancer cells), and in vivo (xenograft mice) preclinical model systems.

Results OLA increased RES-mediated DNA damage, downregulated the HR proteins, caused a late S/G2 arrest, enhanced apoptosis and cell death in RES pre-treated breast cancer cells at much lower concentrations than their individual treatments. Direct measurement of HR pathway activity using a GFP plasmid-based assay demonstrated reduced HR efficiency in I-SceI endonuclease-transfected cells treated with OLA. Moreover, RES+OLA treatment also caused significant reduction in PARP1-mediated PARylation and efficiently trapped PARP1 at the DNA damage site. Upon RES treatment, PARylated PARP1 was found to interact with BRCA1, which then activated other HR proteins. But after addition of OLA in RES pre-treated cells, PARP1 could not interact with BRCA1 due to inhibition of PARylation. This resulted in deregulation of HR pathway. To further confirm the role of BRCA1 in PARP1-mediated HR pathway activation, BRCA1 was knocked down that caused complete inhibition of HR pathway activity, and further enhanced apoptosis after RES+OLA treatment in BRCA1-silenced cells. In agreement with in vitro data, similar experimental results were obtained in ex vivo patient-derived breast cancer cells and in vivo xenograft mice. Conclusion RES+OLA combination treatment enhanced breast cancer cell death by causing excessive DNA damage and also by simultaneously inhibiting the HR pathway.