BACKGROUND:Non-small cell lung cancer (NSCLC) is considered to have more than 80% of all lung cancer cases, making it the leading cause of cancer-related deaths globally. MicroRNA (miRNA) deregulation has been seen often in NSCLC and has been linked to the disease's genesis, progression, and metastasis via affecting their target genes.MATERIALS AND METHODS:Our study focused on the functionality of down-regulated miRNAs in NSCLC. For this study, we used 91 miRNAs reported to be down-regulated in NSCLC. The targets of these miRNAs were chosen from miRNA databases with functionality in NSCLC, including miRBase, miRDB, miRTV, and others. Inter-regulatory miRNA-NSCLC networks were generated. Simulated annealing was used to improve the network's resilience and understandability. GSEA was used to examine 24607 genes reported experimentally in order to gain physiologically relevant information about the target miR-520c-3p.RESULTS:The study revealed functional prominence on miR-520c-3p, down-regulated during NSCLC. The involvement of miR-520c-3p in the PI3K/AKT/mTOR signaling pathway was recognized.CONCLUSIONS:The therapeutic usage by designing a synthetic circuit of miR-520c-3p was explored, which may help in suppressing tumors in NSCLC. Our study holds promise for the successful deployment of currently proposed miRNA-based therapies for malignant disorders, which are still in the early pre-clinical stages of development.

Systems pharmacology helps to understand the complex relationships between biological systems, drugs, and infection model; Leishmania major being one of them. It has aided the drug discovery process by addressing the concerns about economic stress, drug toxicity, and the emergence of resistance. Two million new leishmaniasis cases are reported annually, and >350 million people are at risk globally due to the parasite Leishmania. Try-panothione reductase (TryR) from the parasite-specific redox metabolism is a promising target. In the discipline of medicinal chemistry, benzimidazole is a strong pharmacophore and exhibits a broad range of biological ac-tivities. In the current study, benzimidazole derivatives were explored using computational, enzyme kinetics, biological activity, cytotoxic impact characterization, and in-silico ADME-Tox predictions, followed by their confirmation through in-vitro and animal experiments to discover novel inhibitors for TryR from Leishmania major. During rigorous in-silico screening, two benzimidazole derivatives were chosen for further experimenta-tion. In-vitro testing revealed that compound C1 has a higher binding affinity for the TryR protein. Treatment with compound C1 caused significant morphological changes in the parasite, including size reduction, membrane blebbing, loss of motility, and improved anti-leishmanial efficacy. The compound C1 had significant anti-leishmanial potential against L. major promastigotes and demonstrated apoptosis-mediated leishmanicidal ac-tivity (apoptosis-like cell death). Furthermore, BALB/c female mice treated with C1 reduced parasite burden. Our findings depicts that C1 successfully lowered the parasite load and has a therapeutic impact on infected mice making C1 as a promising lead compound that, with additional modifications, may be exploited to create novel anti-leishmanial therapies.

With the necessity to develop antileishmanial drugs with substrate specificity, trypanothione reductase (TryR) has gained popularity in parasitology. TryR is unique to be present only in trypanosomatids and is functionally similar to glutathione in mammals. It protects against oxidative stress exerted by the host defense mechanism. The TryR enzyme is essential for the survival of Leishmania parasites in the host as it reduces trypanothione and aids in neutralizing hydrogen peroxide produced by the host macrophages during infection. Henceforth, it becomes vital to decipher their functional stability and behaviour in the presence of denaturants. Our study is focused on structural, functional and behavioural stability aspects of TryR with different concentrations of Urea, Guanidinium chloride, alcohol based compounds followed by extensive molecular dynamics simulations in a lipid bilayer system. The results obtained from the study reveal an interesting insight into the possible mechanisms of modulation of the structure, function and stability of the TryR protein.

Cytokines are influential molecules which can direct cells behavior. In this review, cytokines are referred as messengers, immune cells which respond to cytokine stimulus are referred as receivers and the immune cells which gets modulated due to their plasticity induced by infectious pathogen leishmania, are referred as followers. The advantage of plasticity of cells is taken by the parasite to switch them from parasite eliminating form to parasite survival favoring form through a process called as reciprocity which is undergone by cytokines, wherein pro-inflammatory to anti-inflammatory switch occur rendering immune cell population to switch their phenotype. Detailed study of this switch can help in identification of important targets which can help in restoring the phenotype to parasite eliminating form and this can be done through synthetic circuit, finding its wider applicability in therapeutics.

Medicinal plants serve as a rich source of therapeutic modalities against various diseases especially cancer, which gained attention by researchers, due to the problems associated with available treatments. Indian traditional medicinal systems have a rich repository of medicinal plants and could serve as a source of safe and cost-effective alternative therapy. A major limitation with natural products is meager information about their mode of action, hindering their wider clinical use. Herein, we are focusing on anticancer potential of few medicinal plants based on common chemical constituents, viz. Peganum harmala, Quercus infectoria, Melissa officinalis, and Plumbago zeylanica, with well-cited use in folk medicines. Anticancer mechanism of these plants/active components has been well deciphered with targeted pathways involved in their anticancer potential. Peganum harmala and its active constituents showed involvement of various signaling pathways for anticancer effect. Cancer cell death by Quercus infectoria and its constituents also involved pathways such as AKT, NF-κB, and JAK/STAT. Melissa officinalis showed anticancer potential by affecting various transcription factors such as NF-κB, TNF-α, and COX-2. Anticancer activity mediated by plumbagin isolated from Plumbago also showed effect on various transcription factors/signaling pathways as pTEN, mTOR, and Akt pathways, thereby inhibiting survival signaling. However, overall data suggested the need for more extensive studies focused on clinical investigation for pharmacokinetics, bioavailability, and toxicity of these plants/plant products. We propose in this chapter that studies based on computational biology using identified pure compounds and their target to develop pharmacophore model for drug discovery would be a fast and feasible way to design new potential derivatives.

Leishmaniasis, the second most neglected tropical disease, has been reported to affect approximately 12 million people worldwide. The causative protozoan parasite Leishmania has shown drug resistance to available chemotherapies, owing to which we need to look for better approaches to deal with the clinical situations. As per recent reports, several miRNAs have been found to be differentially expressed during Leishmania major infection in host macrophages. We aim to evaluate the impact of miRNA-mediated gene regulation on the key players of inflammation and macrophage dysfunction. The origin of Leishmania miRNAs and their processing is a questionable phenomenon as of yet. Through our study, we aim to provide a framework of their characterization. We amalgamate chemical systems biology and synthetic biology approaches to identify putative miRNA targets and unravel the complexity of host-pathogen gene regulatory networks.

Autophagy is a self-destructing mechanism of cell via lysosomal degradation, which helps to degrade/ destroy hazardous substances, proteins, degenerating organelles and recycling nutrient. It plays an important role in cellular homeostasis and regulates internal environment of cell, moreover, when needed causes non-apoptotic programmed death of cell. It’s usually detailed with ageing and nutritional stress, but recent researches provide support for its role in innate as well as adaptive immunity. Autophagy has been observed as one of the major factors in parasite clearance in leishmaniasis. Due to intra-cellular pathogen, the cell mediated response is only alternative for adaptive immunity against Leishmania in host. T-cells have been observed as main component of cell mediated immunity in leishmaniasis. The differentiation of T-cells generates either destruction or proliferation of parasite as different kind of cytokine mediate different results. Pro-inflammatory cytokine IL12 and TNFα generate Th2 response which helps in active phagocytosis of parasite whereas an anti-inflammatory cytokine like IL10 mediate parasite promotion by blocking autophagic pathways and inhibiting phagocytic actions. TLR2/6 mediated signaling stimulated by LPG produces many pro-inflammatory cytokines like IL12, TNFα and IL6 etc. In macrophages, it is found that TNFα via autocrine signaling induce autophagy and help in parasite killing of Toxoplasma gondii via JNK phosphorylation in macrophages. The phosphorylated JNK induces Beclin by phosphorylating Bcl2, which inhibits Beclin action, thus releasing Beclin and promoting VpS34 which recruits PI3P for autophagosome formation. Recruitment of PI3P is controlled by ATG9 protein, autophagy related gene protein, which in normal condition is inhibited by mTOR via ULK/ ATG1. IL10 has been observed to inhibit autophagy via starvation induced AKT-PI3K pathway in macrophages and result in inducing mTOR. Thus, we can conclude that IL10 inhibits recruitment of PI3P via mTOR. Through systems aspect, here, we decipher that Atg9-PI3P acts as a negative feedback loop in autophagic machinery of leishmaniasis.

Systems and synthetic biology in the coming era has the ability to manipulate, stimulate and engineer cells to counteract the pathogenic immune response. The inherent biological complexities associated with the creation of a device allow capitalizing the biotechnological resources either by simply administering a recombinant cytokine or just reprogramming the immune cells. The strategy outlined, adopted and discussed may mark the beginning with promising therapeutics based on the principles of synthetic immunology.