Novel nanoformulation of Sabizabulin (VERU-111) for pancreatic cancer therapy

Background: Pancreatic cancer (PanCa) is one of the leading causes of cancer-related mortality in the United States due to very limited therapeutic options. Thus, developing novel therapeutic strategies will help for the management of this disease. We recently identified VERU-111, a novel synthetic molecule which showed potent anti-cancer effect against PanCa via targeting clinically important βIII and βIV tubulin isoforms. In this study, we synthesized and characterized its novel nanoformulation (MNP-VERU) and evaluated its therapeutic effects in vitro and xenograft mouse model. Methods: MNPs were prepared by chemical precipitation method and loaded with VERU-111 using diffusion method. This formulation was characterized for particle size, chemical composition, and drug loading efficiency, using various physico-chemical methods (TEM, FT-IR, DSC, TGA, and HPLC). The internalization of MNP-VERU was achieved after 6 hours incubation with MNP-VERU in PanCa cells. To determine therapeutic efficacy of MNP-VERU, we performed various in vitro (MTS, wound healing, boyden chamber real-time xCELLigence, and apoptosis assays) and in vivo (mouse tumor xenograft) studies using PanCa. Effect of MNP-VERU on various key oncogenic signaling pathways, and miRNAs was evaluated by Western blot, immunohistochemistry (IHC), confocal microscopy, qRT-PCR and in situ hybridization (ISH) analyses respectively. Results: Our novel MNP-VERU formulation provided an average size of 110 nm in dynamic light scattering (DLS) and exhibited -8.23 to -11.65 mV zeta potential with an outstanding loading efficiency (94%). Cellular uptake and internalization studies demonstrate that MNP-VERU escapes lysosomal degradation, providing efficient endosomal release to cytosol. MNP-VERU showed remarkable anti-cancer potential in various PanCa cells (Panc-1, AsPC-1, HPAF-II, BxPC-3, MiaPaca) and more effectively repressed βIII and βIV tubulin isoforms via restoring the expression of miR-200c. MNP-VERU more effectively suppressed AsPC-1 cells derived xenograft tumors in athymic nude mice. Conclusions: Taken together, our results suggest that MNP-VERU has more anti-cancer potential than free VERU-111 against PanCa. MNP-VERU may reduce the toxicity and improve the bioavailability of free VERU-111 and could be used for the management of PanCa and health disparity

Smoking and Drinking Activate NF-κB /IL-6 Axis to Promote Inflammation During Cervical Carcinogenesis

Background: High-risk strains of HPV are known to cause cervical cancer. Multiple clinical studies have emphasized that smoking and drinking are critical risk factors for cervical cancer and its high-grade precursors. In this study, we investigated the molecular mechanisms involved in the interplay of smoking and/or drinking with HPV infectivity and defined a systematic therapeutic approach for their attenuation in cervical cancer. Methods: The impact of benzo[a]pyrene (B[a]P) and/or ethanol (EtOH) exposure on cervical cancer cells was assessed by measuring changes in their biophysical, cell migration, and invasion characteristics. Expression of HPV16 E6/E7, NF-κB, cytokines, and inflammation mediators was determined using qRT-PCR, immunoblotting, ELISA, luciferase reporter assay, and confocal microscopy. Results: Treatments with B[a]P and/or EtOH altered the expression of HPV16 E6/E7 oncogenes and EMT markers in cervical cancer cells; it also enhanced migration and invasion. In addition, B[a]P and/or EtOH exposure promoted inflammation pathways through TNF-α and NF-κB signaling, leading to IL-6 upregulation and activation of VEGF. The molecular effects caused by B[a]P and/or EtOH exposure were effectively attenuated by curcumin (Cur)/PLGA-Cur treatment. Conclusions: These data suggest a molecular link between smoking, drinking, and HPV infectivity in cervical carcinogenesis. In addition, attenuation of these effects by treatment with Cur/PLGA-Cur treatment implies the role of curcumin

Proteomics of Extracellular Vesicles: Recent Updates, Challenges and Limitations

Extracellular vesicles (EVs) are lipid-bound vesicles secreted by cells, including exosomes, microvesicles, and apoptotic bodies. Proteomic analyses of EVs, particularly in relation to cancer, reveal specific biomarkers crucial for diagnosis and therapy. However, isolation techniques such as ultracentrifugation, size-exclusion chromatography, and ultrafiltration face challenges regarding purity, contamination, and yield. Contamination from other proteins complicates downstream processing, leading to difficulties in identifying biomarkers and interpreting results. Future research will focus on refining EV characterization for diagnostic and therapeutic applications, improving proteomics tools for greater accuracy, and exploring the use of EVs in drug delivery and regenerative medicine. In this review, we provide a bird’s eye view of various challenges, starting with EV isolation methods, yield, purity, and limitations in the proteome analysis of EVs for identifying protein targets

Physics Potential of the ICAL detector at the India-based Neutrino Observatory (INO)

The upcoming 50 kt magnetized iron calorimeter (ICAL) detector at the India-based Neutrino Observatory (INO) is designed to study the atmospheric neutrinos and antineutrinos separately over a wide range of energies and path lengths. The primary focus of this experiment is to explore the Earth matter effects by observing the energy and zenith angle dependence of the atmospheric neutrinos in the multi-GeV range. This study will be crucial to address some of the outstanding issues in neutrino oscillation physics, including the fundamental issue of neutrino mass hierarchy. In this document, we present the physics potential of the detector as obtained from realistic detector simulations. We describe the simulation framework, the neutrino interactions in the detector, and the expected response of the detector to particles traversing it. The ICAL detector can determine the energy and direction of the muons to a high precision, and in addition, its sensitivity to multi-GeV hadrons increases its physics reach substantially. Its charge identification capability, and hence its ability to distinguish neutrinos from antineutrinos, makes it an efficient detector for determining the neutrino mass hierarchy. In this report, we outline the analyses carried out for the determination of neutrino mass hierarchy and precision measurements of atmospheric neutrino mixing parameters at ICAL, and give the expected physics reach of the detector with 10 years of runtime. We also explore the potential of ICAL for probing new physics scenarios like CPT violation and the presence of magnetic monopoles.Comment: 139 pages, Physics White Paper of the ICAL (INO) Collaboration, Contents identical with the version published in Pramana - J. Physic

Integrative analysis of transcriptomics and clinical data of SLC1A5 in cervical cancer

Background: Cervical cancer is the fourth most common cancer in adult females globally and the second most common cancer in females between the ages of 15 and 44. The alanine-serine- cysteine transporter 2, ASCT2 (solute carrier family 1 member 5, SLC1A5), is a major transporter of the amino acid, glutamine. SLC1A5 expression is associated with the progression of multiple types of tumors, including liver, lung, breast, colon and head and neck cancer. However, the biological significance of SLC1A5 cervical cancer has not been systematically and comprehensively investigated. Using deep data mining and transcriptomics analyses, in this study, we investigated the significance of SLC1A5 expression and its association with cervical cancer prognosis. Methods: SLC1A5 mRNA expression in cervical cancer was examined using the Cancer Genome Atlas (TCGA), the Genotype Tissue Expression Atlas (GTEx), the Tumor Immune Estimation Resource (TIMER), and the UALCAN databases to examine the relationship between the expression of SLC1A5 and the clinicopathological characteristics of cervical cancer. The genetic alterations and frequency of SLC1A5 were analyzed using the cBioPortal and COSMIC databases. The correlations between SLC1A5 and tumor-infiltrating immune cells were examined using the TIMER 2.0, TISIDB, and GEPIA databases. The data processing analysis is based on the R language. LinkedOmics was used for SLC1A5 co-expression network analysis. Drug sensitivity was analyzed through the GDSC and CTRP databases. Results: The SLC1A5 mRNA expression was upregulated in cervical cancer samples as compared to normal cervical tissues. For the cancer stages, ages, races, patients weight, histopathological subtype, and nodal metastasis status there existed significant difference in the expression of SLC1A5 among different groups by mining of the UALCAN database. The mutation landscape analysis confirms that SLC1A5 genetic alterations reached 3%, of which missense substitutions accounted for the highest proportion of 48.77%. The findings of the TIMER analysis indicated a correlation between immune cell infiltration and SLC1A5 copy number alterations (CNA). The expression level of SLC1A5 was positively correlated with the infiltration level of CD4+T cells, Neutrophil, and dendritic cell but negatively correlated with CD8+ T cells, B cells and macrophages infiltration. Additionally, SLC1A5 expression was also found to be associated with certain immunosuppressive cells, chemokines, and receptors. Through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, SLC1A5 expression was found to be closely related to some immune pathways, including proteasome, DNA replication, ribosome biogenesis and DNA replication. Among all co-expressed genes, NOX3 and SHMT2 had a high co-expression efficient. Furthermore, SLC1A5 presented a positive association with sensitivity to GDSC and CTRP drugs. Conclusion: Our observation demonstrates that SLC1A5 was highly expressed in cervical cancer tissues and is linked to prognosis of cervical cancer patients and tumor immune responses. The present study might be benefit for better understanding the clinical significance of SLC1A5 and provided a potential therapeutic target for cervical cancer in future

Nebulization based Inhalation Nanomedicine for Lung Cancer Treatments

Background: Lung cancer is reported to have a high incidence rate and first leading cause of cancer-related morbidity and mortality across the world including in the United States. Noninvasive nebulized inhalation is a promising delivery strategy for lung, which can enhance the targeting efficiency and detention time interval of nanoparticles in the lung tissue, thus elevating the therapeutic index of therapeutic agent(s) at lower dosages. The aim of this study is to develop inhalable nanoparticles (INPs) for effective delivery of therapeutic agents in lung cancer cell lines and ex vivo models. Methods: The inhalation nanoparticles (INPs) were prepared by solvent evaporation and self-assembly approach. The INPs formulations were characterized by particle size, chemical composition, and drug loading efficiency using various analytical methods including FT-IR, DSC, SEM, and DSC/TGA. Cellular uptake of INPs was evaluated in 2D and 3D models of lung cancer cell lines (A549 and NCI-H1299) using fluorescence microscopy and flow cytometry analysis. Additionally, the therapeutic evaluation of gambogic acid and gemcitabine encapsulated INPs was performed by basic in vitro biological assays using proliferation (CCK-8), mucoadhesion Boyden chamber, and apoptosis assays using lung cancer (A549 and NCI-H1299) monolayers, spheroids, and xenograft tumors. Results: The developed INPs exhibited an average size of ~110 nm in dynamic light scattering measurements. INPs formulation showed a remarkable mucoadhesion and mucopenetration potential in-vitro model(s). Cellular uptake studies demonstrated that INPs formulation facilitates an effective endosomal release into the cytosol. The in vitro study confirms that INPs release the drugs in a sustained manner. Additionally, the INPs formulation showed superior in vitro anti-cancer activity in lung cancer cell lines, spheroids and xenograft tumor. Conclusions: Altogether this study confirms that INPs formulation demonstrates an improved therapeutic benefit over free drug against lung cancer cell lines, spheroids and xenograft tumor. This study could lead as an innovative therapeutic modality for the treatment of lung cancer

Withania somnifera: Progress towards a Pharmaceutical Agent for Immunomodulation and Cancer Therapeutics

Chemotherapy is one of the prime treatment options for cancer. However, the key issues with traditional chemotherapy are recurrence of cancer, development of resistance to chemotherapeutic agents, affordability, late-stage detection, serious health consequences, and inaccessibility. Hence, there is an urgent need to find innovative and cost-effective therapies that can target multiple gene products with minimal adverse reactions. Natural phytochemicals originating from plants constitute a significant proportion of the possible therapeutic agents. In this article, we reviewed the advances and the potential of Withania somnifera (WS) as an anticancer and immunomodulatory molecule. Several preclinical studies have shown the potential of WS to prevent or slow the progression of cancer originating from various organs such as the liver, cervix, breast, brain, colon, skin, lung, and prostate. WS extracts act via various pathways and provide optimum effectiveness against drug resistance in cancer. However, stability, bioavailability, and target specificity are major obstacles in combination therapy and have limited their application. The novel nanotechnology approaches enable solubility, stability, absorption, protection from premature degradation in the body, and increased circulation time and invariably results in a high differential uptake efficiency in the phytochemical\u27s target cells. The present review primarily emphasizes the insights of WS source, chemistry, and the molecular pathways involved in tumor regression, as well as developments achieved in the delivery of WS for cancer therapy using nanotechnology. This review substantiates WS as a potential immunomodulatory, anticancer, and chemopreventive agent and highlights its potential use in cancer treatment

Confounding Factors of Cervical Carcinogenesis and Cervical Cancer Health Disparity

Clinical studies show that cervical cancer is the most frequently detected cancer in women living with human immunodeficiency virus (HIV), which has been defined as an acquired immunodeficiency syndrome (AIDS). Women living with HIV are 6 times more likely to develop cervical cancer compared to the general population, and an estimated 5% of all cervical cancer cases are attributable to HIV. Studies suggest that several genes are involved in different molecular pathways, which can influence the vulnerability, severity, or advancement of cervical cancer in the presence of HIV and human papillomavirus (HPV) co-infection. Clinical studies have also shown levels of cervical COX-2 are increased in HIV-infected and HIV/HPV-coinfected women. However, the precise molecular-level interactions between HPV and HIV co-infection are not well defined. Further, smoking is well documented among HIV-positive women, but very little is known about the smoking-induced molecular mechanism of HPV-HIV co-infections. In this study, we investigated the interplay between smoking and HIV/HPV co-infection infectivity and their cooperativity. To check the influence of HIV-HPV co-infection at the molecular level, cervical cancer cells, CaSki, were treated with U1(HIV-1-infected U937 cells) cell culture supernatant (UCS) and U937 (HIV-1 noninfected cells) culture supernatant (U9CS) every day for 4 days. To differentiate the U1 cells into macrophages, U1 cells were treated with 100 nM phorbol 12-myristate 13-acetate (PMA) for 3 days. For the treatment of smoking components, both U937 and U1 cells were treated with benzo[a]pyrene (B[a]P) at 100 nM every 24 hours for 4 days. Similarly, we treated HIV-infected (U1) and noninfected (U937) cells were treated with HPV-infected (CasKi) cells supernatant to determine if HPV infection can influence HIV replication-related molecular pathways. Our study demonstrated that HIV-HPV-co-infection and/or B[a]P enhanced cell proliferation colony formation, migration, and invasion as demonstrated by MTS, wound healing, and Boyden chamber assay, respectively, in CaSki cells. HIV-HPV-co-infection and/or B[a]P enhanced also induced the expression of HPV16 E6/E7, HIF-1α, PTGS2, and EMT markers in CaSki cells as determined by qRT-PCR and immunoblotting. Furthermore, our qRT-PCR results demonstrated that cytokine IL-6, IL-8, IP-10, MCP-1, IL-1α, IL-10, MIP-1β TNF-α, IFN-γ and G-CSF were significantly increased in CaSki cell treated with UCS and B[a]P. We have also demonstrated that the supernatant of cervical cancer cells exacerbates HIV-1 replication in differentiated U1 cell lines via transferring CYPs and HPV oncoproteins through extracellular vehicles. Taken together, our results demonstrate that a molecular link between smoking and HIV infections in HPV cooperativity in cervical carcinogenesis might also be responsible for cervical cancer disparity among underserved populations

Antibody-Drug Conjugates for Cancer Therapy: Chemistry to Clinical Implications

Chemotherapy is one of themajor therapeutic options for cancer treatment. Chemotherapy is often associated with a low therapeutic window due to its poor specificity towards tumor cells/tissues. Antibody-drug conjugate (ADC) technology may provide a potentially new therapeutic solution for cancer treatment. ADC technology uses an antibody-mediated delivery of cytotoxic drugs to the tumors in a targeted manner, while sparing normal cells. Such a targeted approach can improve the tumor-to-normal tissue selectivity and specificity in chemotherapy. Considering its importance in cancer treatment, we aim to review recent efforts for the design and development of ADCs. ADCs are mainly composed of an antibody, a cytotoxic payload, and a linker, which can offer selectivity against tumors, anti-cancer activity, and stability in systemic circulation. Therefore, we have reviewed recent updates and principal considerations behind ADC designs, which are not only based on the identification of target antigen, cytotoxic drug, and linker, but also on the drug-linker chemistry and conjugation site at the antibody. Our review focuses on site-specific conjugation methods for producing homogenous ADCs with constant drug-antibody ratio (DAR) in order to tackle several drawbacks that exists in conventional conjugation methods

Integrative transcriptomics data analysis of TRPV1 in hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) is the most common type of liver cancer. Transient receptor potential vanilloid 1 (TRPV1) is a nonselective Ca2+ channel protein that is widely expressed and plays a significant role in cancer initiation and progression. However, the biological significance of TRPV1 in HCC has not been systematically and comprehensively investigated. Using deep data mining and transcriptomics analyses, in this study, we described the significance of TRPV1 expression and its association with HCC prognosis. Methods: TRPV1 mRNA expression in HCC was examined using the Cancer Genome Atlas (TCGA), the Genotype Tissue Expression Atlas (GTEx), the Tumor Immune Estimation Resource (TIMER), and the UALCAN databases to examine the relationship between the expression of TRPV1 and the clinicopathological characteristics of HCC. The genetic alterations and frequency of TRPV1 were analyzed using the cBioPortal and COSMIC databases. The correlations between TRPV1 and tumor-infiltrating immune cells were examined using the TIMER 2.0, TISIDB, and GEPIA databases. The data processing analysis is based on the R language. LinkedOmics was used for TRPV1 co-expression network analysis. Results: TRPV1 mRNA expression was upregulated in HCC samples as compared to normal liver tissues. Kaplan-Meier analysis demonstrated that high expression of TRPV1 is associated with better prognostic significance for overall survival (OS) and disease-free survival (DFS) in HCC patients. The mutation landscape analysis confirms that TRPV1 genetic alterations reached 6%, of which missense substitutions accounted for the highest proportion of 16.16%. The findings of the TIMER analysis indicated a correlation between immune cell infiltration and TRPV1 copy number alterations (CNA). The expression level of TRPV1 was positively correlated with the infiltration level of CD4+ T cells but negatively correlated with CD8+ T cells, B cells, macrophages, and dendritic cell infiltration. Additionally, TRPV1 expression was also found to be associated with certain immunosuppressive cells, chemokines, and receptors. Through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, TRPV1 expression was found to be closely related to some immune pathways, including drug metabolism, PPAR signaling pathway, and chemical carcinogenesis. Conclusion: Our observation demonstrates that TRPV1 was highly expressed in HCC tissues and is linked to prognosis of HCC patients and tumor immune responses