Nilesh Kumar Sharma

Breast cancer stem cells as last soldiers eluding therapeutic burn: A hard nut to crack

Cancer stem cells (CSCs) are found in many cancer types, including breast carcinoma. Breast cancer stem cells (BCSCs) are considered as seed of cancer formation and they are associated with metastasis and genotoxic drug resistance. Several studies highlighted the presence of BCSCs in tumor microenvironment and they are accentuated with several carcinoma events including metastasis and resistance to genotoxic drugs and they also rebound after genotoxic burn. Stemness properties of a small population of cells in carcinoma have provided clues regarding the role of tumor microenvironment in tumor pathophysiology. Hence, insights in cancer stem cell biology with respect to molecular signaling, genetics and epigenetic behavior of CSCs have been used to modulate tumor drug resistance due to genotoxic drugs and signaling protein inhibitors. This review summarizes major scientific breakthroughs in understanding the contribution of BCSCs towards tumors capability to endure destruction inflicted by molecular as well as genotoxic drugs.

Export of microRNAs: A Bridge between Breast Carcinoma and Their Neighboring Cells

Breast cancer is a leading type of cancer among women in India as well as worldwide. According to the WHO 2015 report, it has been anticipated that there would be a twofold rise in the death due to breast cancer among women. The heterogeneous property of breast carcinoma has been suggested to be linked with dedicated set of communication and signaling pathway with their surroundings, which culminate into progression and development of the cancer. Among the plethora of communication tools in the hand of breast carcinoma cells is the recently appreciated exocytosis of the tightly packed short non-coding RNA molecules, predominantly the microRNAs (miRNAs). Recent studies suggest that miRNAs may work as courier messengers to participate in endocrine and paracrine signaling to facilitate information transfer between breast carcinoma and their neighboring cells. Evidence suggests that breast tumor cells communicate via packaged miRNAs in the tumor-released microvesicles, which enrich the tumor microenvironment. There is a strong view that dissecting out the mechanistic and regulatory aspects of miRNA export and role may uncover many prospects for overcoming the signaling defects and thereby controlling aberrant cell division. The detection of circulating miRNAs associated with breast carcinoma can also be used as biomarkers for early diagnosis. This review article is an attempt to provide updated knowledge on implications of short RNAs and their transport in the breast cancer pathophysiology.

Aberrant DNA Double-strand Break Repair Threads in Breast Carcinoma: Orchestrating Genomic Insult Survival.

Breast carcinoma is a heterogeneous disease that has exhibited rapid resistance to treatment in the last decade. Depending genotype and phenotype of breast cancer, there are discernible differences in DNA repair protein responses including DNA double strand break repair. It is a fact that different molecular sub-types of breast carcinoma activate these dedicated protein pathways in a distinct manner. The DNA double-strand damage repair machinery is manipulated by breast carcinoma to selectively repair the damage or insults inflicted by the genotoxic effects of chemotherapy or radiation therapy. The two DNA double-strand break repair pathways employed by breast carcinoma are homologous recombination and non-homologous end joining. In recent decades, therapeutic interventions targeting one or more factors involved in repairing DNA double-strand breaks inflicted by chemo/radiation therapy have been widely studied. Herein, this review paper summarizes the recent evidence and ongoing clinical trials citing potential therapeutic combinatorial interventions targeting DNA double-strand break repair pathways in breast carcinoma.

Mutual concessions and compromises between stromal cells and cancer cells: driving tumor development and drug resistance

BackgroundVarious cancers have been found to be associated with heterogeneous and adaptive tumor microenvironments (TMEs) and to be driven by the local TMEs in which they thrive. Cancer heterogeneity plays an important role in tumor cell survival, progression and drug resistance. The diverse cellular components of the TME may include cancer-associated fibroblasts, adipocytes, pericytes, mesenchymal stem cells, endothelial cells, lymphocytes and other immune cells. These components may support tumor development through the secretion of growth factors, evasion from immune checkpoints, metabolic adaptations, modulations of the extracellular matrix, activation of oncogenes and the acquisition of drug resistance. Here, we will address recent advances in our understanding of the molecular mechanisms underlying stromal-tumor cell interactions, with special emphasis on basic and pre-clinical information that may facilitate the design of novel personalized cancer therapies.ConclusionsThis review presents a holistic view on the translational potential of the interplay between stromal cells and cancer cells. This interplay is currently being employed for the development of promising preclinical and clinical biomarkers, and the design of small molecule inhibitors, antibodies and small RNAs for (combinatorial) cancer treatment options. In addition, nano-carriers, tissue scaffolds and 3-D based matrices are being developed to precisely and safely deliver these compounds.

Modulating secreted components of tumor microenvironment: A masterstroke in tumor therapeutics

ABSTRACT The microenvironment in which cancer resides plays an important role in regulating cancer survival, progression, malignancy and drug resistance. Tumor microenvironment (TME) consists of heterogeneous number and types of cellular and non-cellular components that vary in relation to tumor phenotype and genotype. In recent, non-cellular secreted components of microenvironmental heterogeneity have been suggested to contain various growth factors, cytokines, RNA, DNA, metabolites, structural matrix and matricellular proteins. These non-cellular components have been indicated to orchestrate numerous ways to support cancer survival and progression by providing metabolites, energy, growth signals, evading immune surveillance, drug resistance environment, metastatic and angiogenesis cues. Thus, switching action from pro-cancer to anti-cancer activities of these secreted components of TME has been considered as a new avenue in cancer therapeutics and drug resistance. In this report, we summarize the recent pre-clinical and clinical evidences to emphasize the importance of non-cellular components of TME in achieving precision therapeutics and biomarker study.

Epigenetic perturbation driving asleep telomerase reverse transcriptase: Possible therapeutic avenues in carcinoma

In the last decade, implications of human telomerase reverse transcriptase (hTERT), a component of ribonucleoprotein telomerase in aging, senescence, and stem cell are highly evident. Besides, the activation of hTERT is also being documented several cancer types including carcinoma. The awakening of telomerase during carcinoma initiation and development is being seen with different perspectives including genetic and epigenetic tools and events. In view of several tumor progenitors genes (also referred as epigenetic mediators), telomerase is placed as key enzyme to achieve the carcinoma phenotype and sustain during the progression. It is true that swaying of telomerase in carcinoma could be facilitated with dedicated set of epigenetic modulators and modifiers players. These epigenetic alterations are heritable, potentially reversible, and seen as the epigenetic signature of carcinoma. Several papers converge to suggest that DNA methylation, histone modification, and small non-coding RNAs are the widely appreciated epigenetic changes towards hTERT modulation. In this review, we summarize the contribution of epigenetic factors in the telomerase activation and discuss potential avenues to achieve therapeutic intervention in carcinoma.

DNA Repair Response Modulation Potentiates Low Dose Cisplatin Effects in HeLa Cells

Background: Cisplatin is considered as the crucial regimen of widely prescribed chemotherapy treatment to many cancer types. However, incessant increasing dose of cisplatin against many cancers including cervical cancer is known for drug resistance and side effects like nephrotoxicity

Beyond gene dictation in oral squamous cell carcinoma progression and its therapeutic implications

In recent, mechanisms behind transition from normal epithelium to premalignancy and finally to oral carcinoma have been linked to genetic and epigenetic multistep process. These epigenetic changes are heritable, potentially reversible, and presented as epigenetic signature normal and carcinoma cells. Such epigenetic alterations have been suggested as DNA methylation, histone modifications, and small noncoding RNAs. These extragenetic changes have been reported to play a role in the modulation of several key signaling, regulatory, and repair routes in oral squamous cell carcinoma (OSCC) and other carcinomas. In this review, we summarize scientific developments in the last decade, substantiating the role of extra- and beyond genetic molecular signatures in OSCC progression, and potential biomarker and implications in chemotherapy intervention.

Epigenomic hard drive (EHD) imprinting: A hidden code within cancer cell to survive beyond the biological death of a tumor patient

L are fructose polymers synthesized by a broad range of microorganisms and a limited number of plant species as nonstructural storage carbohydrates and they have potential applications in the pharmaceutical, food, and cosmetic industries. The present study shows a comparative analyses of polysaccharide type Levan biosynthesis in static and shaking fermentation by using Z. mobilis ATCC 10988 strain. All fermentation processes were carried out in Erlenmeyer flasks presenting a capacity of 500 ml and working volume of 100 ml culture medium, in both fermentation types: static and on the rotary shaker, with stirring of 220 rpm, with the temperature maintenance at 32°C for 72 hours. In figure 1, we can find the results obtained by static fermentation for different initial concentrations of sucrose. The best development of the microbial culture in terms of microbial biomass was seen for the initial concentration of 15% sucrose, due probably to the favorable ratio between the source of C and N. The largest amount of polysaccharide was obtained from the experiment with 40% initial sucrose (8.9 g%), but this value is also similar to the experiment with 25% initial sucrose (8.47 g%). The Figure No. 2 shows the results obtained by the stirred fermentation with the strain Z. mobilis ATCC 10988 for different initial concentrations of sucrose. Concerning the content of polysaccharide, if the initial concentration of sucrose in the biosynthesis medium was greater than 20%, the amount of polysaccharide was about 5 g%, without any increase of production in the case of higher concentrations. By comparing the results shown in figures 1 and 2, it can be noticed that the strain Z. mobilis ATCC 10988 performs better concerning the polysaccharide biosynthesis in the frame of a static fermentation, which is not a surprise considering that these bacteria are an optional aerobic one.S rufopilosa, a tsema rowan, is a species of the small ornamental trees in the genus Sorbus and the family Rosaceae found in East Asia. The antioxidant and anticancer effects of S. rufopilosa remain unclear. The objective of this study is to evaluate the antioxidant and anticancer effects of ethanol extract of S. rufopilosa (EESR) and the molecular mechanism of its anticancer activity in human colon carcinoma HT29 cells. EESR showed significant antioxidant activity and inhibitory effect on HT29 cell growth in a dose-dependent manner. EESR induced cell cycle arrest at G2/M phase in a dose-dependent manner by modulating the expression of cyclin B, cyclin-dependent kinase 1 (CDK1), and CDK inhibitor p21. EESR-induced apoptosis was associated with the upregulation of p53, a death receptor Fas, a pro-apoptotic protein Bax and the activation of caspase 3, 8, and 9, resulting in the degradation of poly ADP ribose polymerase (PARP). These results suggest that EESR efficiently inhibits proliferation of HT29 by inducing both cell cycle arrest and apoptosis, and may be a possible candidate for the anticancer drug development. Recent Publications: 1. Andreas G (2003) Introduction to apoptosis. ApoReview 2-26. 2. Evan GI, Vousden KH (2001) Proliferation, cell cycle and apoptosis in cancer. Nature 411:342-348. 3. Fulda S (2015) Targeting apoptosis for anticancer therapy. Semin. Cancer Biol. 31:84-88. 4. Fulda S, Debatin KM (2006) Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 25:4798-4811. 5. Udensi UK, Tchounwou PB (2016) Oxidative stress in prostate hyperplasia and carcinogenesis. J. Exp. Clin. Cancer Res. 35:139.I clinical drug development, it is important to understand the absorption, distribution, metabolism and excretion (ADME) properties of a drug in humans. The micro-tracer study based on the accelerator mass spectrometry (AMS) is an ultrasensitive technique to obtain human ADME profiles with a negligible radiation dose. KD101 is a novel compound under development to treat obesity. The aim of this study was to investigate the absorption, metabolism and excretion properties of KD101 in obese subjects. A randomized, open-label, single-dose, one-treatment, one-period, one-sequence study was conducted in six males with a BMI ≥27, who received KD101 at 400 mg with 3.52 μg of [14C]-KD101 (180 nCi) in the fed state. Plasma, urine and feces samples were collected up to 288 hours post-dose for mass balance and metabolite profiling. Plasma concentrations of KD101 were determined using a validated GC method. Total radioactivity in the samples was determined using AMS. Safety and tolerability was evaluated based on vital signs, adverse events, clinical laboratory tests, and electrocardiography. All of the subjects completed the study with no clinically significant safety issue. Mean total recovery rate (range) was 85.21% (75.3699.01%), consisting of 77.96% (68.31-92.33 %) for urine and 7.26% (5.91-8.51%) for feces, which differed greatly from the preclinical data. Oral absorption of [14C]-KD101 was rapid with the peak plasma concentration reaching at 5.83h post dose, which was consistent with the previous report. In the urine radiochromatogram, five large peaks were identified including a peak represented by the parent compound. KD101 is excreted predominantly through the urine in humans. Many of the excreted materials in the urine were considered metabolites. This study demonstrated effectiveness of the micro-tracer study enabled by AMS in humans to investigate the ADME property of KD101, which hugely differed from that seen in the preclinical animals.Objectives: YH4808 is a highly potent, selective and reversible potassium-competitive acid blocker (P-CAB) on H/K-ATPase under development for the treatment of gastric acid related diseases including gastroesophageal reflux disease and peptide ulcer disease. The objectives of this study were 1) to develop a human PBPK model optimized by human pharmacokinetic (PK) data and 2) to predict the PK profiles of YH4808 using the PBPK model in various clinical settings.Statement of the Problem: High performance liquid chromatography (HPLC, including UHPLC) is one of the most powerful separation techniques capable of providing analysis difficult or impossible with other separation approaches. Thus, chromatographers give particular importance to the design of new efficient stationary phases for HPLC. New trends in chromatographic separations have been directed towards the use of multi-modal stationary phases (MM-SPs) which are high resolution, high selectivity, high loading capacity, high speed, minimal solvent consumption compared with single-modal stationary phases (SM-SPs).Aim: To compare the serum levels of adiponectin, vitamin D, copper and zinc in rheumatoid arthritis (RA) patients and to investigate the relationship between these factors and disease severity. Method: Ninety patients with RA and 30 healthy individuals were participated in this study according to the ACR/ EULAR criteria for RA. Serum concentrations of adiponectin were determined by ELISA, copper and zinc by colorimetric spectrophotometry and vitamin D by HPLC methods. Results: Serum adiponectin and vitamin D were increased and decreased in RA patients, respectively. Adiponectin and disease severity are positively correlated, whereas vitamin D and disease severity are negatively correlated. Adiponectin negatively correlate with vitamin D and positively correlated with disease activity score (DAS). Copper and zinc showed no significant difference between two groups. Conclusion: The definitive roles of adiponectin, vitamin D, copper and zinc is not completely determined. More investigations are needed to deeply explore the impact of them on RA pathophysiology. Finally, we suggest these serum factors as promising diagnostic and therapeutic biomarkers.C stem cells (CSCs) possess characteristics associated with normal stem cells and may generate tumors through the stem cell processes of self-renewal and differentiation into multiple cell types. They involved in drug resistance, metastasis and relapse of cancers can significantly affect tumor therapy. Therefore, it is important to develop specific therapies targeted probe at CSCs for improvement of survival and quality of life of cancer patients. Studies have indicated that the CD166 protein has been considered as a specific marker for colorectal CSCs detection. In addition to monoclonal antibodies, small molecules such as anti-peptides could provide more advantages for CSCs detection in vivo. Here, we attended to design the CD166 antipeptide (CD166ap) to detecting the CSCs in vitro and in vivo. To obtain CSCs, the human colorectal cancer cells (HCT-15) were seeded into selection media (DMEM/F12, 0.4% bovine serum albumin, 2% B27, 5 μg/mL bovine insulin, 4 μg/mL heparin, 20 ng/mL fibroblast growth factor 2, and 20 ng/mL epidermal growth factor) at a density of 1000 cells per mL. Under these conditions, only CSCs and early progenitor cells survive and proliferate, whereas differentiated cells die. Next, we designed a CD166ap (amino acid: KDSEGYESYNGNLGSQC {It is known that human CD6 proteins have the ability to bind the human CD166 proteins specifically (Chappell PE et al., Structure. 2015, 23:1426-1436). Depending on the two proteins binding sites, we designed the amino acid sequence of CD166 anti-peptide. However, the Nand C-terminal amino acid (lysine and cysteine) were added for conjugating with fluorescence, nuclear medicine chelator and Polyethylene glycol (PEG).} and conjugated with fluorescence for CSCs binding assay by flow cytometry and immunofluorescence stain. For in vivo imaging detection, the media-induced CSCs (2×106) were subcutaneous inoculated into the right flank of nude mice (n=5 per each group) and grew for one week. Then, the fluorescence conjugated CD166ap was IV injected into animal model for in vivo imaging system and biodistribution assay. The primary spheres that derived from HCT-15 cells under serum-free conditions and which are highly enriched for CSCs at 48 hours. These induced CSCs overexpressed the reprogramming factors such as OCT4, c-myc, Nanog and anti-apoptosis factor (Survivin). Moreover, they also showed the characteristic of drug resistance compared with cancer cells. In CSCs targeted probe binding assay, the CD166ap and antibody revealed the quite binding capability in CSCs. The in vivo imaging assay, we found that CD166ap specifically targeted to CSCs-induced xenograft model and accumulated in tumor area. In conclusion, we designed a specific probe for CSCs detection in vivo successfully. In addition, the CD166ap may label radioisotope for nuclear medicine imaging and conjugate drug for CSCs therapy in clinical.L (LBL) coating has gained popularity for drug delivery of therapeutic drugs. Herein, we described an approach for enhancing the therapeutic efficiency of the locally administered dexamethasone (Dx) for the treatment of inflammatory bowel disease (IBD). We utilized a LBL-coating technique for alternative coating of Dx microcrystals (DxMCs) with multiple layers of polyelectrolytes composed of poly (allylamine hydrochloride), poly (sodium 4-styrene sulfonate) and Eudragit® S100. The successful deposition of the layers onto DxMCs surfaces were confirmed through zeta potential measurement and confocal laser scanning microscopy, while the surface morphology was investigated through scanning electron microscopy. The drug encapsulation efficiency for LBL-DxMCs was 95% with a mean particle size of 2 μm and negative surface charge of -45 mV. Moreover, in vitro drug release studies showed a minimum release of the drug ( 15%) at an acidic condition during initial first 5 h followed by sustained-release at alkaline condition. For in vivo study, LBL-DxMCs were administered orally to male ICR mice suffering from dextran sulfate sodium-induced colitis. LBL-DxMCs was found to substantially enhance antiinflammatory efficacy of the drug compared to uncoated DxMCs. Macroscopic, histological and biochemical (tumor necrosis factor-α, interleukin-6 and myeloperoxidase) examinations revealed marked improvements of colitis signs in the mice treated with LBL-DxMCs compared with those treated with uncoated DxMCs. Overall, the obtained results demonstrate that LBLDxMCs are an effective and safe colon-targeted delivery system for the treatment of inflammatory bowel disease.Results: Seven RCTs were found in the systematic review (639 patients) and were included in the meta-analysis. There is not clinical difference in the parasitological cure between MBZ and metronidazole (MTZ). The relative risk (RR) is 0.88 (95% CI 0.70-1.10), with a moderate heterogeneity (I2=66%). The prediction interval expected to cover the results of a new trial is wide enough (0.35-2.21) to support both a parasitological relevant difference favoring MBZ and a parasitological relevant difference favoring MTZ.L are fructose polymers synthesized by a broad range of microorganisms and a limited number of plant species as nonstructural storage carbohydrates and they have potential applications in the pharmaceutical, food, and cosmetic industries (1-5). This study presents a comparative analysis of the growth and consumption curves of Z. mobilis NCIB 1163 and Z. mobilis ATCC 10988, levan producing microorganisms. The growth and consumption curves were performed in a liquid medium with a concentration of 5% sucrose and 5% glucose. Thus, the bacterial cells in the exponential growth phase were centrifuged at 12.000 g and the pellet was washed twice with a sterile 0.9% NaCl solution. Finally, the cells were resuspended in 1 ml of physiological saline and they were used as inoculum in 5% liquid medium, which was monitored concerning the fermentable carbohydrate consumption and the growth. The consumption curves were performed under anaerobic conditions (the culture medium was coated with a layer of sterile paraffin oil) by periodical weighing of the medium seeded with different bacterial strains of Z. mobilis and by the graphical representation of weight loss (due to release of carbon dioxide). Exponentially growing cells were used as the inoculum, made at approximately 107 cells/ml. Cell growth was assayed turbidimetrically at a wavelength of 600 nm. The consumption curves of Z. mobilis NCIB 11163 and Z. mobilis ATCC 10988 bacterial strains on complete sucrose substrate medium, 5% under anaerobic conditions (Figure 1) revealed that strains NCIB 11163 and ATCC 10988 exhibit a similarly kinetics of consumptions substrate. The consumption curves of Z. mobilis NCIB 11163 and Z. mobilis ATCC 10988 bacterial strains on complete glucose substrate medium, 5% under anaerobic conditions (Figure 2) show that glycolysis is more intense than hydrolysis of sucrose. From the curves profile it is observed that the strain ATCC 10988 shows a curve having a more pronounced linear decrease.

An evidence of black box within cancer patient: epigenomic hard drive (EHD) imprinting

* Hadeer A Abbassy 1 , Reham A Aboelwafa 1 and Omar M Ghallab 2 . 1. Clinical pathology department, Faculty of Medicine, Alexandria University, Egypt. 2. Clinical hematology department, Faculty of Medicine, Alexandria University, Egypt. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History