ABSTRACT

Medulloblastomas are the most common brain tumor in childhood that originates from cerebellar granule cell precursors (GCPs), located in the external granular layer (EGL) of the cerebellum. The antiproliferative gene PC3 promotes cerebellar neurogenesis by inducing GCPs to shift from proliferation to differentiation. There are crossed transgenic mice conditionally expressing PC3 in GCPs with Patched1 heterozygous mice (Ptc +/-), a model of medulloblastoma pathogenesis is characterized by hyperactivation of the Sonic Hedgehog pathway. Up-regulation of PC3 in Ptc +/- mice results in a decrease of medulloblastoma incidence of ~40% and in  a marked reduction of preneoplastic abnormalities, such as hyperplastic EGL areas and lesions. Overexpression of cyclin D1, hyperproliferation, and defective differentiation, observed in Ptc +/- GCPs, are restored to normality in Ptc +/- mice. The PC3-mediated inhibition of cyclin D1 expression correlates with recruitment of PC3 to the cyclin D1 promoter, which is accompanied by histone deacetylation. Remarkably, down-regulation of PC3 is observed in preneoplastic lesions, as well as in human and murine medulloblastomas. As a whole, this indicates the PC3 may prevent medulloblastoma development by controlling cell cycle and promoting differentiation of GCPs.

ABSTRACT

Acute myeloid leukemia (AML) is a clinically and genetically heterogeneous disease characterized by aberrant proliferation and differentiation of myeloid progenitor cells that affect patients of all ages. Despite current advances in understanding AML pathophysiology and improved overall outcomes, the standard treatment regimen has remained unchanged over several decades and ineffective for specific AML subtypes. The U.S. Food and Drug Administration (FDA) has approved several targeted therapeutics for AML subtypes with different genetic aberrations in recent years. The complex cytogenetics of AML is also associated with a higher relapse rate and leads to a poor prognosis. So, there is an urgent need for new therapeutic agents which can substantiate and supplement the existing therapies for AML treatment. As the process of conventional drug development suffers from high cost and low success rate, alternative strategies such as drug repurposing have gained tremendous attention in recent years. Drug repurposing or repositioning is an approach to identify new therapeutic applications for existing drugs that are approved with well-known toxicity profiles and reasonable prices. Since the medications for repurposing use need to be validated directly in a clinical setting, drug repositing offers a time- and cost-effective approach for identifying new therapies compared to traditional drug development. Several FDA-approved non-chemotherapy drugs have been under investigation in recent years for their efficacy against AML treatment. As AML is a highly heterogeneous disease, identification of additional agents with a potential to exhibit antileukemic activity through a drug repurposing approach would augment existing AML therapies. Therefore, the drug repurposing approach might offer better cancer therapeutics for AML treatment, and their efficacy in combination with standard of care therapy would be of potential interest in the future for treating hematological malignancies.

ABSTRACT

As one of the  speakers in this Conference on cancer research and therapy, I will just outline a brief overview of my personal journey from basic science research in USA to translational breast cancer research in Chennai, India.

This journey involved value teaching principles implemented in hometown Chennai in 2000 in  Biotechnology , Molecular Biology and Immunology , all of which was important to make an early transition to Translational medicine research in breast cancer as early as 2005-2007,when the scientific World had not woken up to the tremendous volume  of inhibitors in targeted therapy churned out, ,over 2000 in breast cancer alone, and only a few showing promising results in clinics back then. Metastasis and relapse were not taking centre stage back then. Preclinical animal models were not sufficiently adequate to mimic human picture.

We  took up the study  of breast cancer diagnostic ( biomarkers)  and breast cancer therapeutic s in photothermal abalation of breast cancer tumors, with the view to have startup or small scale industry to implement the same. But this road less travelled upon , required a thorough awareness of the culture we were embedded in, even in a University setting, which we did not possess till now, and litigation became necessary to blow the hot breezes of unscientific and unrelated bureaucracy, so that the light once lit can be passed on to the younger, if they dare to hold it. That’s the romance of study of oncology – the study of biomarkers that pinpoint the subset of patients that would benefit from the treatment given and research of therapeutic s that would propel us out of the rut of 2-3 month or even year of benefit, to complete elimination of the disease itself.

ABSTRACT

Ovary is one of the most important organ in female reproductive system. It performs two main functions: secretion of oocytes with which when the sperm cells fuse fertilization occurs, and the secretion of female sex hormones estrogen and progesterone which are important for the expression of feminine characteristics in female. Ovary has three main compartments: the outer epithelial layer, next inner layer is cortex and the innermost cells are medulla [1,2]. The main organelles which perform the above mentioned vital functions of ovary are ovarian follicles which crowd within the cortex. Given these basic heterogeneities in the structure of ovary and that within the structure of overall female reproductive system and abdomen, the ovarian cancer is highly heterogeneous and is of many types: epithelial ovarian carcinoma (serous, endometrioid, mucinous and clear cell), germ cell tumor, stromal cell tumor [3]. The origins of these ovarian cancers have long been in doubt and several hypotheses have been proposed regarding these. None of these hypotheses have been proven and the confusion remains. Taking advantage of the fact that in the last 10-15 years both tissue-specific stem cells have been discovered in healthy ovary [4] and Ovarian Cancer Stem Cells have been discovered in cancerous ovary [3], I try my best to address the confusions over the origins of ovarian cancers.  

ABSTRACT

To improve effective localised delivery and drug retention during biological discharge, this research targeted intravesical Gemcitabine (GTB) delivery of cationic polymeric nanoparticles to treat ovarian cancer. Initially, four GTB loaded polymer nanoparticles, i.e., chitosan nanoparticles (CS-NPs), polysarcosin nanoparticles (PSar-NPs), poly-llysine & polysarcosin nanoparticles (PLL-PSar-NPs), chitosan & polysarcosin nanoparticles (CS-PSar-NPs), were prepared. Based on preliminary particle size, zeta potential, encapsulation efficiency, DSC, surface morphology, release profiling, cellular internalization studies using Rhodamine123 & Nile red fluorescent markers, it was hypothesised that CS-PSar-NPs may be the best cationic formulation with strong biocompatibility and anti-cancer activity against OVCAR-8 ovarian cancer cell line. Epidermal growth factor receptor variant III (EGFR vIII) anchored over all four polymeric nanoparticles to increase successful targeting, better cellular penetration and in-vitro cytotoxicity. It was verified from confocal microscopy that EGFRvIII conjugated cationic GTB polymeric nanoparticles were showing higher cellular uptake with double internalization capability than unconjugated nanoparticles with time-dependent cell entry. Harboring GTB and EGFRvIII combined polymer nanoparticles would have a greater ability to penetrate ovarian cancer cells during the initial incubation hour. TEM findings suggest that EGFRvIII conjugation over the non-target CS-PSar-NP surface was successful, making CS-PSar-NPS-EGFRvIII more target-specific and presumably a safer candidate for ovarian cancer.

ABSTRACT

Present investigation demonstrated the cancer treatment depends on numerous molecular functions and mechanisms that promote tumour growth through the immune system. Immunologic theorem suggested cancer immunotherapy based on molecular immune checkpoints controls T-cell activation. Those mechanisms foster novel therapeutic applications in Immuno-Oncology. So, the FASLG gene from the TNF family binds with the receptor to induce apoptosis. Fas ligand and receptor interactions lead to a dominant nature in the immune system and develop cancer. The dynamism of the apoptotic-induced FASLG gene terminates tumour infiltrating lymphocytes and suppress tumour responses called tumour counter-attacks. Intense evidence in the animal model suggested FASLG (CD95L) gene expression prevent T-cells responses and tumours exit. The justification of CD95L (CD178) in tumours decline and induce inflammation. Those mechanisms suggested CD178 in tumour cells and its contribution to immune break. In this work, I aimed to a genome-wide survey of the TNF family to pursue the molecular immunologic mechanisms linked with the FASLG gene in the mammalian genome. So, I performed bioinformatics and computational applications to gain awareness of the FASLG gene and its family in two different organisms. A genome-wide observation suggested the numeral of the FASLG gene and TNF domains in Homo sapiens and Mus musculus. Therefore, I observed the FASLG and their functional mechanisms integrated with immune response. Those mechanisms forwarded the T-cell suppression in the tumour micro-environment by immune checkpoints controlling tumours growth to interfere with anti-tumour immune response.

ABSTRACT

In approximately 23% of AML patients, Fms-like tyrosine kinase 3 (FLT3) contains a gain-of-function, internal tandem duplication (ITD) mutation that is associated with an unfavourable prognosis. FLT3 is therefore seen as a promising therapeutic avenue for AML and as a result a more complete understanding of its function and signalling may lead to additional targets and resistance mechanisms being identified. To date, several FLT3 inhibitors including Quizartinib (AC220), a second-generation ITD-selective tyrosine kinase inhibitor, have been developed, but clinical trials when used as single agents have not been overwhelmingly successful. In most preclinical studies, the inhibitory effects of FLT3 inhibitors are mainly evaluated using mutant -expressing models, however, most AML patients harbour both a wild-type and mutant FLT3 allele as well as presenting with high plasma levels of FLT3 ligand (FLT3L). We hypothesized that FLT3L could act to influence the efficacy of FLT3 inhibitors through WT-FLT3 in cells with heterozygous mutations. In this study, we have examined the role of FLT3 inhibition and FLT3L on the cellular localisation and downstream signalling in several AML cell lines, some of which express ITD mutations. We also looked at mechanisms by which FLT3L could impair the efficacy of FLT3 inhibitors. Our results revealed that the majority of FLT3 in the MV4-11 and MOLM-13 cells was intracellular. Inhibition of ITD-FLT3 with quizartinib led to a dramatic relocalization of FLT3L to the cell surface. This effect was more pronounced in MV4-11 (FLT3ITD/ITD) than the heterozygous MOLM-13 (FLT3ITD/WT) cells which also express WT. This was accompanied by inhibition of ERK, AKT and STAT5 signaling pathways and resulted in cell death.  Quizartinib induced cell death only in AML lines expressing FLT3-ITD mutations (MOLM-13 and MV4-11), and this could be antagonised by FLT3 ligand (FLT3L). The largest inhibitory effects were seen in heterozygous cells (those expressing a mutated and wild-type allele). FLT3 inhibition was associated with downregulation of the anti-apoptotic Mcl-1 and upregulation of the pro-apoptotic BH3-only protein, BIM. siRNA experiments indicated that both of these proteins were regulated by both WT and ITD-FLT3 through the MAPK pathway. These results suggest that activation of FLT3 signalling by FLT3L confers chemoresistance to quizartinib through suppression of BIM expression and upregulation of MCL-1. Taken together, our data suggest that cell surface localization, FLT3 and FLT3L expression are controlled by ITD mutations and are key components of drug resistance. The data also suggests a novel therapeutic approach in patients with high plasma FLT3L levels, especially when using type II inhibitors such as quizartinib, is to co-target the MAPK pathway to allow BIM expression.  
 

ABSTRACT

The interaction between TSGs and other genes in DNA leads to the development of cancer. TSGs can be turned off, lost, or rendered ineffective (loss of heterozygozity) due to genetic alterations other than mutations in the DNA sequence. Cancer prevention, diagnosis, and treatment are all impacted by epigenetic silencing. In cancer cells, drugs that repair epigenetic changes and restore gene activity have now been approved. Furthermore, because changes in DNA methylation may be detected with high sensitivity, numerous approaches that check for changes in DNA methylation can be used to detect cancer early. Early research has suggested that CAR T-cell therapy for GBM is viable, safe, and may even be therapeutic. The challenges ahead include optimizing infusion dosage and frequency, regulating the immunosuppressive tumor microenvironment, and dealing with the significant genetic variation. CARs (chimeric antigen receptors) are recombinant receptors made up of an extracellular antigen-recognition moiety, which is usually generated from an antibody. T cells that have been transplanted with CARs gain the ability to identify and lyse cancer cells. CARs have the ability to detect TAs in a variety of forms, including carbohydrates, glycolipids, and proteins. T cell receptors detect mainly intracellular components that are delivered by antigens, whereas CARs require the target TA to be produced on the cell membrane. By inhibiting the inhibitory receptor programmed death 1 (PD-1), which is produced by T cells, immune resistance can be overcome. For anticancer efficacy and safety, an antibody that specifically targets PD-1 was considered. Long-term tumor reduction and disease stability were observed in patients with antibody-mediated PD-L1 suppression. CAR T cells are a highly promising therapy for GBM with many potential target TAs identified. Selectivity and heterogeneity of a suitable TA, especially one that will lead to the elimination of GBM CICs, represents one of the current major challenges in the field. There is still a long way ahead for CAR T cell therapy before it becomes a standard of care for the treatment of patients with GBM. Antibodies that inhibit one or both of these receptors suggest that antitumor immunity can be improved and should provide a promising strategy of immunotherapy for selected tumors expressing PD-L. 
 

ABSTRACT

The T regulatory cells (Tregs) are major TME infiltrating immune cells. The Tregs that express FoxP3 shield tumor cells from the antitumor immune response by regulating cofactor and functional pathways. The tumor microenvironment (TME) of triple-negative breast cancer (TNBC) is highly immunogenic and poorly responds to immunotherapeutics due to the enrichment of FoxP3+Tregs. The reduced antitumor immune response contributes to poor prognosis as well as a high risk of recurrence. Therefore, understanding the biology of TME infiltrating Tregs is essential to evaluate diagnostic and prognostic relevance in envisaging response to therapy. This review discusses the role of FoxP3 and its partners in the regulation of immune checkpoints in Tregs. This review highlighted the functional pathways of FoxP3+Tregs and mechanisms that regulate immune-suppressive functions in TME. We also highlighted strategies of therapeutic blockade of FoxP3 in BC. Finally, we reviewed the prognostic relevance of TME infiltrating Tregs in TNBC to envisage response to therapy. The central theme of this review is to highlight the diagnostic and prognostic potential of TME infiltrating Tregs in TNBC.

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Introduction: Some risks factors assotiated wuth COVID 19 infections were estableished such as coronary artery disease in patients with SARS-CoV-2 infections. The use of highly concentrated medications accorfing to Ardnt Shultz Law can induce reduction in the vital energy, leading to stagnation of Blood and propensity to have myocardail infaction even without arterail obstruction. Purpose; to demonstrate that patient with COVID 19 infection that received highly concentrated medications to treat this infection, have more propensity to  develop myocardila infarction some days after the treatment instituted. Methods; through one case report of 42 years-old patient with history of acquired COVID 19 on january 2nd 2021, he was admited in the hospital due to dyspnea symptons, myalgia, and needs oxigenation. He received the medications, ivermectine, hydroxicloroquine, corticosteroids . After 18 days of the initial manifestations, the patient felt pain in the chest and went to the hospital and they found that his troponin were increasing gradually and after two hours of it increased many times and the physician decided to admit him again in the hospital to make more exams. The patient was submitted to cattheterization and the result of this procedure were that all his coronary were in perfect state of health without any obstruction and they treated him as he had myocardial infaction. After three months , the patient went to the author´s clinic to evaluate his condition and she did chakras’ energy centers measurement.  Results: the results of this condition  that revealed that all his chakras’ were in the lowest level of energy with exception of the seventh that was normal. The author began his treatment with the use og homeopathies medications according to the theory Constitutional Homeopathy of the Five Elements based on Traditional Chinese Medicine. Conclusion; the conclusiopn of this study is that patients when treated with highlt concentrated medications to treat SARS-CoV-2 infection can develop myocadial ingaction without arterial obstruction due to energy deficiency state that agraveted many times due to the use of highly concentrrated medications used to treat this kind of infection nowadays.                  

ABSTRACT

Humans are exposed to various free radical generating pro-oxidative stressors such as environmental pollutants and carcinogen, which modulate the host immune response to diseases, including cancer. One of the consequences of these prooxidative stressors is an exaggerated production of a potent lipid mediator, Platelet-activating factor (PAF) via reactive oxygen species (ROS). In asmuch as most therapeutic agents also generate ROS as one of the primary mechanisms for inducing cytotoxicity leading to tumor cell death, our studies determined the functional significance and translational relevance of these oxidized PAF agonists in the efficacy of cancer therapies. Once generated, these PAF agonists mediate their effects via binding to a G-protein coupled, PAF-receptor (PAFR), expressed on various cell types, including tumor cells. Using PAFR-expressing and deficient cellular systems, mouse models, and pharmacological agents, our studies have demonstrated that clinically-relevant chemotherapeutic agents and radiation therapy all produce PAF agonists. These oxidized PAF agonists are blocked by antioxidants, PAF-metabolizing PAF-acetyl hydrolase (PAF-AH) enzyme, inhibitors of cyclooxygenase type 2 (COX-2) enzyme, and PAFR antagonists. Notably, in mouse models, chemotherapy, or radiation therapy treatment of one tumor was found to increase the growth of secondary (non-treated) tumors, indicating an abscopal effect of oxidized PAF agonists in supporting tumor growth, as well as limiting the efficacy of therapeutic agents. Importantly, increased PAFR activity and tumoral PAF were detected in the perfusates/tumor samples of cancer patients isolated post-treatment compared to the pre-treatment. Mechanistically, we have discovered that metabolically-labile PAF agonists travel via small membrane-shed bioactive vesicles known as microvesicle particles (MVP), which mediate the therapeutic agents-generated PAF agonists responses, in a process blocked by inhibitors of PAFR and MVP biogenesis enzyme. Overall, our studies provide compelling evidence of PAFR signaling in impacting the efficacy of therapeutic agents and device approaches to circumvent these PAF agonists-mediated effects to improve therapy effectiveness.  

ABSTRACT

Studies have demonstrated that cooking of meat at high temperature such as frying or barbeque causes production of heterocyclic amines (HCAs). At least a dozen of HCAs are found in cooked meat. The 2-amino-1-methyl-6-phenylimidazo[4-5-b]pyridine (PhIP) is abundant and most potent HCA in cooked meat. Several studies have shown that PhIP can induce tumors in breast, prostate and colon cells and in rodent models. It is shown that PhIP causes DNA mutation, promote tumor growth and promote invasiveness of cancer cells. PhIP metabolites are known to produce DNA adduct and DNA strand breaks. Phytonutrients are known to inhibit cytotoxic and genotoxic effects. Therefore, we hypothesized that the right combination of phytonutrients (naturally present in fruits, vegetables and spices) along with grilled meat should be capable of suppressing the PhIP induced cytotoxic actions.  

Therefore, a model system using human breast epithelial cells (MCF 10A) was developed to test and understand the epigenetic interaction of PhIP and phytonutrients. We tested several phytonutrients to evaluate their benefits from PhIP. Results indicate that presence of phytonutrients improves cell viability as compared to cells treated only with PhIP. Curcumin found in turmeric seems to be the best in protecting MCF 10A cells from the PhIP induced carcinogenicity. Multiple analysis were performed to evaluate interactions include - cell viability assay, DCF assay to quantify ROS production, Comet assay to quantify the DNA damage and DNA adduct formation by immunofluorescence procedure.  Curcumin co-treated cells showed significant differences and PhIP induced cell cytotoxicity was consistently relapsed to normal. Gene expression studies showed that curcumin is capable of interacting via multiple molecular targets. Therefore, our results suggest that curcumin appears to be an effective anti-PhIP food beneficial additive. There is an urgent need to educate people about preventive measures contributing to our health.

ABSTRACT

Liquid biopsies offer a promising alternative to tissue samples, providing non-invasive diagnostic approaches or serial monitoring of disease evolution. However, certain challenges remain, and the full potential of liquid biopsies has yet to be reached. Here we report several methodological approaches to interrogate liquid biopsies using circulating tumour cell (CTC) enumeration and characterisation, transcriptomics, circulating free DNA (cfDNA) isolation and copy number instability (CNI) scores using blood samples from lung, ovarian and anal cancer patients.  Collectively or data suggests that obtaining reliable readouts from blood can provide crucial information for disease progression, as well as being of prognostic value, monitoring patients’ response to treatment.