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PDF file - 433KB, S1: CIP2A and SET levels are increased in some primary human pancreatic cancer samples. S2: Increased expression of PP2A inhibitors trend toward decreased PP2A activity, which trends toward increased Myc expression and Myc protein stabilization in pancreatic cancer cell lines. S3: OP449 treatment decreases c-Myc DNA binding and half-life in pancreatic cancer cell lines, and SET knockdown reduces OP449 sensitivity. S4: OP449 treatment reduces pancreatic cancer cell migration in vitro, proliferation in vivo, and increases tumor cell apoptosis in vivo.

Seminal discoveries have established that epigenetic modifications are important for driving tumor progression. Polycomb group (PcG) proteins are highly conserved epigenetic effectors that maintain, by posttranslational modification of histones, the silenced state of genes involved in critical biologic processes, including cellular development, stem cell plasticity, and tumor progression. PcG proteins are found in two multimeric protein complexes called Polycomb repressive complexes: PRC1 and PRC2. Enhancer of zeste homolog 2 (EZH2), catalytic core subunit of PRC2, epigenetically silences several tumor-suppressor genes by catalyzing the trimethylation of histone H3 at lysine 27, which serves as a docking site for DNA methyltransferases and histone deacetylases. Evidence suggests that overexpression of EZH2 is strongly associated with cancer progression and poor outcome in disparate cancers, including hematologic and epithelial malignancies. The regulatory circuit and molecular cues causing EZH2 deregulation vary in different cancer types. Therefore, this review provides a comprehensive overview on the oncogenic role of EZH2 during tumorigenesis and highlights the multifaceted role of EZH2, as either a transcriptional activator or repressor depending on the cellular context. Additional insight is provided on the recent understanding of the causes and consequences of EZH2 overexpression in specific cancer types. Finally, evidence is discussed on how EZH2 has emerged as a promising target in anticancer therapy and the prospects for targeting EZH2 without affecting global methylation status. Thus, a better understanding of the complex epigenetic regulatory network controlling EZH2 expression and target genes facilitates the design of novel therapeutic interventions. Mol Cancer Res; 12(5); 639–53. ©2014 AACR.

R-loops, or RNA:DNA hybrids, can induce DNA damage, which requires DNA repair factors including breast cancer type 1 susceptibility protein (BRCA1) to restore genomic integrity. To date, several pathogenic mutations have been found within the tandem BRCA1 carboxyl-terminal (BRCT) domains that mediate BRCA1 interactions with proteins and DNA in response to DNA damage. Here, we describe a nonrepair role of BRCA1 BRCT in suppressing ribosomal R-loops via two mechanisms. Through its RNA binding and annealing activities, BRCA1 BRCT facilitates the formation of double-stranded RNA between ribosomal RNA (rRNA) and antisense-rRNA (as-rRNA), hereby minimizing rRNA hybridization to ribosomal DNA to form R-loops. BRCA1 BRCT also promotes RNA polymerase I-dependent transcription of as-rRNA to enhance double-stranded rRNA (ds-rRNA) formation. In addition, BRCA1 BRCT-mediated as-rRNA production restricts rRNA maturation in unperturbed cells. Hence, impairing as-rRNA transcription and ds-rRNA formation due to BRCA1 BRCT deficiency deregulates rRNA processing and increases ribosomal R-loops and DNA breaks. Our results link ribosomal biogenesis dysfunction to BRCA1-associated genomic instability.

Dry eye disease (DED) is a multifactorial disorder of the ocular surface that results in ocular discomfort, visual disturbance and damage to the ocular surface. It is one of the most common complaints in daily ophthalmic practice. The greater prevalence of dry eye in women compared to men suggests that sex hormones may have a role in this condition. Sex hormones; estrogen and androgens influence production of all components of the tear film including aqueous layer, lipid layer, and mucin layer. Various mechanisms such as decrease in hormonal levels, shift in feedback mechanisms, and changes in receptor receptivity interplay to alter the ocular surface homeostasis and subsequently result in DED. The purpose of this review is to briefly outline current scientific evidence on the influence of androgen and estrogen on the lacrimal and meibomian glands as well as on the ocular surface epithelia including conjunctival goblet cells during reproductive and menopausal periods. This article also outlines the updates regarding role of gonadal hormones in the treatment of dry eye. Keywords: Dry Eye, Sex hormones, Estrogen, Androgen, Testosterone, Progesterone, Tear film, Meibomian gland, Lacrimal gland, Ocular surface, Menopause, HRT, Schirmer’s test.

The study was conducted at the College of Agriculture and Research Station, Indira Gandhi Krishi Vishwavidylaya, Janjgir- Champa (Chhattisgarh), during the year 2017-18 Rabi Season to Study the Influence of Rhizobium and phosphate solubilizing bacteria on the performance of field-grown Lathyrus (Lathyrus sativus L) under climatic conditions of Chhattisgarh plains. In the present investigation total of 8 treatments T1 -Control (No inoculum no fertilizer), T2- Recommended Dose of Fertilizer RDF(N:P: K::20:40:20), T3- Rhi.HG-25 (Std.) + PSB-H-27 (Std.), T4 Rhi.HG-25 + Korba PSB-118, T5 - Rhi.L-3 Lathyrus + PSB-H-27, T6 Rhi.L-3 Lathyrus + Korba PSB-118, T7- Rhi.L-11 Lathyrus + PSB-H-27 , T8 Rhi.L-11 Lathyrus + Korba PSB-118. Recommended Dose of Fertilizer (RDF) T2 recorded maximum and significantly higher value of plant height over rest of the treatments at 45, 60 DAS and at maturity except T7 (Rhi.L-11 Lathyrus + PSB-H-27) treatment which was statistically similar at maturity. A general perusal of the data of the fresh plant reveals that T2 (Recommended Dose of Fertilizer, RDF) recorded the maximum and significantly higher value of fresh weight over the rest of the treatments at a later stage of crop growth i.e 60 DAS and at harvest. A critical perusal of the nodule count data revealed that treatment T8 (Rhi.L-11Lathyrus+ KorbaPSB-118) was found superior over all the treatments showing the superiority of combined use of local strains over other strains and RDF. In case of grain yield the maximum grain yield were observed in 1451.39 kg/ha due to application of T8 (Rhi.L-11Lathyrus+ KorbaPSB-118) followed by T2 RDF 1415.28 kg/ha, T4 (Rhi.HG-25 +KorbaPSB-118)1388.89 kg/ha, T7 (Rhi.L-11 Lathyrus+ PSB-H-27)1344.44 kg/ha, T6 (Rhi.L-3 Lathyrus­ + PSB-H-27)706.94 kg/ha, 12.77 kg/ha, T5 (Rhi.L-3 Lathyrus+ PSB-H-27) 1216.67 kg/ha, T3 (Rhi.HG-25 + PSB-H-27) 1219.94kg/ha. Maximum grain yield was observed due to the application of effective local strains of rhizobia and PSB which also enhanced the nodulation activity due to an increase in the available soil phosphorus. The maximum and significantly higher value of grain yield straw yield (1118.06 kg/ha) was observed due to treatment T8 (Rhi.L-11 Lathyrus+ Korba-PSB-118) which was statistically superior over all the treatments except T2 RDF.

The ADP-ribosylhydrolase ARH3 plays a key role in DNA damage repair, digesting poly(ADP-ribose) and removing ADP-ribose from serine residues of the substrates. Specific inhibitors that selectively target ARH3 would be a useful tool to examine DNA damage repair, as well as a possible strategy for tumor suppression. However, efforts to date have not identified any suitable compounds. Here, we used in silico and biochemistry screening to search for ARH3 inhibitors. We discovered a small molecule compound named ARH3 inhibitor 26 (AI26) as, to our knowledge, the first ARH3 inhibitor. AI26 binds to the catalytic pocket of ARH3 and inhibits the enzymatic activity of ARH3 with an estimated IC(50) of ∼2.41 μm in vitro. Moreover, hydrolysis of DNA damage–induced ADP-ribosylation was clearly inhibited when cells were pretreated with AI26, leading to defects in DNA damage repair. In addition, tumor cells with DNA damage repair defects were hypersensitive to AI26 treatment, as well as combinations of AI26 and other DNA-damaging agents such as camptothecin and doxorubicin. Collectively, these results reveal not only a chemical probe to study ARH3-mediated DNA damage repair but also a chemotherapeutic strategy for tumor suppression.

In this work, we establish Ramanujan–Mordell type formulas associated to certain quadratic forms of discriminants 20k or 32k. In the end, a remark is given to illustrate potential extensions to other related cases.

Drug resistance is one of the trademark features of Cancer Stem Cells (CSCs). We and others have recently shown that paucity of functional death receptors (DR4/5) on the cell surface of tumour cells is one of the major reasons for drug resistance, but their involvement in the context of in CSCs is poorly understood. By harnessing CSC specific cytotoxic function of salinomycin, we discovered a critical role of epigenetic modulator EZH2 in regulating the expression of DRs in colon CSCs. Our unbiased proteome profiler array approach followed by ChIP analysis of salinomycin treated cells indicated that the expression of DRs, especially DR4 is epigenetically repressed in colon CSCs. Concurrently, EZH2 knockdown demonstrated increased expression of DR4/DR5, significant reduction of CSC phenotypes such as spheroid formation in-vitro and tumorigenic potential in-vivo in colon cancer. TCGA data analysis of human colon cancer clinical samples shows strong inverse correlation between EZH2 and DR4. Taken together, this study provides an insight about epigenetic regulation of DR4 in colon CSCs and advocates that drug-resistant colon cancer can be therapeutically targeted by combining TRAIL and small molecule EZH2 inhibitors.

Ten-eleven translocation (TET) family enzymes (TET1, TET2, and TET3) oxidize 5-methylcytosine (5mC) and generate 5-hydroxymethylcytosine (5hmC) marks on the genome. Each TET protein also interacts with specific binding partners and partly plays their role independent of catalytic activity. Although the basic role of TET enzymes is well established now, the molecular mechanism and specific contribution of their catalytic and noncatalytic domains remain elusive. Here, by combining in silico and biochemical screening strategy, we have identified a small molecule compound, C35, as a first-in-class TET inhibitor that specifically blocks their catalytic activities. Using this inhibitor, we explored the enzymatic function of TET proteins during somatic cell reprogramming. Interestingly, we found that C35-mediated TET inactivation increased the efficiency of somatic cell programming without affecting TET complexes. Using high-throughput mRNA sequencing, we found that by targeting 5hmC repressive marks in the promoter regions, C35-mediated TET inhibition activates the transcription of the BMP-SMAD-ID signaling pathway, which may be responsible for promoting somatic cell reprogramming. These results suggest that C35 is an important tool for inducing somatic cell reprogramming, as well as for dissecting the other biological functions of TET enzymatic activities without affecting their other nonenzymatic roles.

Nucleosomal histones are barriers to the DNA repair process particularly at DNA double-strand breaks (DSBs). However, the molecular mechanism by which these histone barriers are removed from the sites of DNA damage remains elusive. Here, we have generated a single specific inducible DSB in the cells and systematically examined the histone removal process at the DNA lesion. We found that histone removal occurred immediately following DNA damage and could extend up to a range of few kilobases from the lesion. To examine the molecular mechanism underlying DNA damage-induced histone removal, we screened histone modifications and found that histone ADP-ribosylation was associated with histone removal at DNA lesions. PARP inhibitor treatment suppressed the immediate histone eviction at DNA lesions. Moreover, we examined histone chaperones and found that the FACT complex recognized ADP-ribosylated histones and mediated the removal of histones in response to DNA damage. Taken together, our results reveal a pathway that regulates early histone barrier removal at DNA lesions. It may also explain the mechanism by which PARP inhibitor regulates early DNA damage repair.

Triple-Negative Breast Cancer (TNBC) has a poor prognosis and adverse clinical outcomes among all breast cancer subtypes as there is no available targeted therapy. Overexpression of Enhancer of zeste homolog 2 (EZH2) has been shown to correlate with TNBC’s poor prognosis, but the contribution of EZH2 catalytic (H3K27me3) versus non-catalytic EZH2 (NC-EZH2) function in TNBC progression remains elusive. We reveal that selective hyper-activation of functional EZH2 (H3K27me3) over NC-EZH2 alters TNBC metastatic landscape and fosters its peritoneal metastasis, particularly splenic. Instead of H3K27me3-mediated repression of gene expression; here, it promotes KRT14 transcription by attenuating binding of repressor SP1 to its promoter. Further, KRT14 loss significantly reduces TNBC migration, invasion, and peritoneal metastasis. Consistently, human TNBC metastasis displays positive correlation between H3K27me3 and KRT14 levels. Finally, EZH2 knockdown or H3K27me3 inhibition by EPZ6438 reduces TNBC peritoneal metastasis. Altogether, our preclinical findings suggest a rationale for targeting TNBC with EZH2 inhibitors.

Chemokine receptor CXCR4 overexpression in solid tumors has been strongly associated with poor prognosis and adverse clinical outcome. However, blockade of CXCL12-CXCR4 signaling axis by inhibitors like Nox-A12, FDA approved CXCR4 inhibitor drug AMD3100 have shown limited clinical success in cancer treatment. Therefore, exclusive contribution of CXCR4-CXCL12 signaling in pro-tumorigenic function is questionable. In our pursuit to understand the impact of chemokine signaling in carcinogenesis, we reveal that instead of CXCR4-CXCL12 signaling, presence of CXCR4 intracellular protein augments paclitaxel resistance and pro-tumorigenic functions. In search of pro-apoptotic mechanisms for CXCR4 mediated drug resistance; we discover that DR5 is a new selective target of CXCR4 in breast and colon cancer. Further, we detect that CXCR4 directs the differential recruitment of transcription factors p53 and YY1 to the promoter of DR5 in course of its transcriptional repression. Remarkably, inhibiting CXCR4-ligand-mediated signals completely fails to block the above phenotype. Overexpression of different mutant versions of CXCR4 lacking signal transduction capabilities also result in marked downregulation of DR5 expression in colon cancer indeed confirms the reverse relationship between DR5 and intracellular CXCR4 protein expression. Irrespective of CXCR4 surface expression, by utilizing stable gain and loss of function approaches, we observe that intracellular CXCR4 protein selectively resists and sensitizes colon cancer cells against paclitaxel therapy in vitro and in vivo. Finally, performing TCGA data mining and using human breast cancer patient samples, we demonstrate that expression of CXCR4 and DR5 are inversely regulated. Together, our data suggest that targeting CXCR4 intracellular protein may be critical to dampen the pro-tumorigenic functions of CXCR4.

Triple Negative Breast Cancer (TNBC) is known to have poor prognosis and adverse clinical outcome among all breast cancer subtypes due to the absence of available targeted therapy for it. Emerging literature indicates that epigenetic reprogramming is now appreciated as a driving force for TNBC pathophysiology. High expression of epigenetic modulator EZH2 (Enhancer of zeste homolog 2) has been shown to correlate with TNBC poor prognosis but the contribution of EZH2 catalytic (H3K27me3) versus non-catalytic EZH2 (NC-EZH2) function in TNBC growth and progression remains elusive. In the process of dissecting the impact of H3K27me3 versus NC-EZH2 function in TNBC pathogenesis, we reveal that selective hyperactivation of H3K27me3 over NC-EZH2 not only promotes TNBC metastasis but also alters the metastatic landscape of TNBC. Using extensive in- vivo live animal imaging, we present conclusive evidence that peritoneal metastasis, particularly splenic metastasis of TNBC is governed by H3K27me3. Transcriptome analyses of hyperactive H3K27me3 cells lead us to discover Cytokeratin-14 (KRT14) as a new target of H3K27me3. Unlike classical H3K27me3 mediated suppression of gene expression, here; we observe that H3K27me3 enhances KRT14 transcription by attenuating the binding of transcriptional repressor Sp1 to its promoter. Further, loss of KRT14 significantly reduces TNBC migration, invasion and splenic metastasis. Finally, genetic ablation of EZH2 or pharmacological inhibition of EZH2 catalytic function by FDA approved drug tazemetostat (EPZ6438) robustly inhibits TNBC peritoneal metastasis. Altogether, our preclinical findings posit a rational insight for the clinical development of H3K27me3 inhibitor like tazemetostat as a targeted therapy against TNBC.

Chronic Myelomonocytic Leukemia (CMML) is a clonal hematologic disorder of older individuals characterized by monocytosis and increased risk of progression to acute myeloid leukemia (AML). Although CMML is characterized by relatively low genetic complexity consistent with clonal expansion, loss-of-function mutations in ten-eleven translocation 2 (TET2) protein are frequent and reported in almost 60% of CMML cases. Consistent with this finding, deletion of the Tet2 gene in mice leads to CMML-like disease. TET2 restrains inflammatory signals through various means, for instance by directly suppressing the expression of the IL6 gene in mouse myeloid cells by recruiting histone deacetylases. Conversely, the levels of pre-inflammatory cytokines including IL6 correlate with pre-leukemic proliferation of Tet2-deficient mouse monocytes and other myeloid cells, suggesting that Tet2 deficiency increases IL-6 expression and increased IL-6 expression in turn drives the proliferation of Tet2-deficient myeloid-lineage cells in a positive feedback loop. This scenario may operate in CMML patients, who show increased levels of inflammatory cytokines and increased expression of interferon-induced genes compared to age-matched healthy individuals. To gain insight into the connection between inflammation caused by TET deficiency and monocyte hyperproliferation, we set up cultures of bone marrow derived macrophages (BMDMs) from inducible TET deficient mice. Using this model, we found that TET-deficient BMDMs show increased proliferation compared to wild type controls; moreover, LPS stimulation leads to significantly increased expression of IL6 and IL1b in TET-deficient BMDMs. Our previous RNA sequencing of expanded TET-deficient myeloid cells in vivo showed that TET deficiency correlated with increased expression of transposable elements (TEs) and induced a DNA damage response, leading us to hypothesize that TET deficiency and increased TE expression may activate DNA and RNA sensing pathways and induce sterile inflammation. I am currently using several molecular approaches to delineate the potential connection of TET deficiency with endogenous inflammation in CMML. There is growing evidence that TET deficiency or non-specific inhibition of TET enzymes in CD8 T cells can potentiate anti-tumor responses. I am now asking whether the TET deficiency-inflammation axis I observe in BMDMs might also enhance anti-tumor responses by increasing proinflammatory responses and hence T cell recruitment and anti-tumor activity. Citation Format: Anup Kumar Singh, Atsushi Onodera, Hugo Sepulveda, Carlos Angel, Anjana Rao. Connecting TET2 deficiency with inflammation in CMML and anti-tumor responses [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6320.

Endocrine therapies are standard-of-care treatments for estrogen receptor (ER) positive breast cancer. However, patients with ER+ breast cancer develop resistance to these therapies and most relapsed patients die with endocrine-resistant metastatic disease. Here we show that resistance to the ER degrader, fulvestrant, is accompanied by epigenetic activation of the transcriptional co-activator VGLL1. Rewiring of the epigenome in therapy resistant cells also results in increased binding of the transcription factor TEAD4. Through interaction with TEAD4, VGLL1 induces the expression of genes implicated in cell proliferation in the resistant cells. We demonstrate that VGLL1 is necessary for the growth of fulvestrant-resistant breast cancer cells. Pharmacological disruption of VGLL1/TEAD4 interaction blocked growth of fulvestrant-resistant cells, accompanied by inhibition of VGLL1/TEAD transcriptional programmes. Furthermore, we identify EGFR as an important downstream VGLL1 target, whereby VGLL1-directed EGFR upregulation sensitises fulvestrant-resistant breast cancer cells to EGFR inhibitors. Taken together, our findings identify VGLL1 as a key transcriptional driver in endocrine-resistant breast cancer and identify new therapeutic approaches for advanced breast cancer patients.

Drug resistance is one of the trademark features of Cancer Stem Cells (CSCs). We and others have recently shown that paucity of functional death receptors (DR4/5) on the cell surface of tumor cells is one of the major reasons for drug resistance, but their involvement in the context of in CSCs is poorly understood. By harnessing CSC specific cytotoxic function of salinomycin, we discovered a critical role of epigenetic modulator EZH2 in regulating the expression of DRs in colon CSCs. Our unbiased proteome profiler array approach followed by ChIP analysis of salinomycin treated cells indicated that the expression of DRs, especially DR4 is epigenetically repressed in colon CSCs. Concurrently, EZH2 knockdown demonstrated increased expression of DR4/DR5, significant reduction of CSC phenotype such as spheroid formation in-vitro and tumorigenic potential in-vivo in colon cancer. TCGA data analysis of human colon cancer clinical samples shows strong inverse correlation between EZH2 and DR4. Taken together, this study provides an insight about epigenetic regulation of DR4 in colon CSCs and advocates that drug resistant colon cancer can be therapeutically targeted by combining TRAIL and small molecule EZH2 inhibitors.

An attempt was made to develop auto-flush machine for cleaning of toilet area/elimination area. To study the auto-flush system, a total of 10 weaners were divided into two groups (having 5 each), viz. G1 (weaners kept in a modified intensive pen with auto flush system) and G2 (weaners in intensive pen without auto-flush). The defecation pattern/trend was observed 24 h consecutively for 5 days. Each weaner defecated 8.40±0.23 and 8.92±0.13 times per day in G1 and G2 respectively. The auto-flush was assessed indirectly based on parameters like hygiene score of floor, hygiene score of animals, amount of water consumed and manpower required for cleaning of pen in 5 days. Hygiene score of floor was higher in G1 (143±0.63) compare to G2 (112.2±1.7). Hygiene score of animals was higher in G1 (5) compared to G2 (2.6). Water consumed for cleaning was lower in G1 (162.8±9.06 litres) than G2 (212±1.4 liters). Manpower requirement in G1 (7.85±0.24 sec) was lower than G2 (175.48±2.11 sec). It can be concluded that all weaners used the elimination area for defecation in G1. The auto-flush system helped in saving water, labour time and maintained the cleanliness of both floor and animal.

In this paper, we find formulas for the number of representations of certain diagonal octonary quadratic forms with coefficients $1,2,3,4$ and $6$. We obtain these formulas by constructing explicit bases of the space of modular forms of weight $4$ on $\Gamma_0(48)$ with character., Comment: 43 pages, 10 Tables, accepted for publication in IJNT

Molecular hybridization of different pharmacophores to tackle both tumor growth and metastasis by a single molecular entity can be very effective and unique if the hybrid product shows drug-like properties. Here, we report synthesis and discovery of a novel small-molecule inhibitor of PP2A–β-catenin signaling that limits both in vivo tumor growth and metastasis. Our molecular hybridization approach resulted in cancer cell selectivity and improved drug-like properties of the molecule. Inhibiting PP2A and β-catenin interaction by selectively engaging PR55α-binding site, our most potent small-molecule inhibitor diminished the expression of active β-catenin and its target proteins c-Myc and Cyclin D1. Furthermore, it promotes robust E-cadherin upregulation on the cell surface and increases β-catenin–E-Cadherin association, which may prevent dissemination of metastatic cells. Altogether, we report synthesis and mechanistic insight of a novel drug-like molecule to differentially target β-catenin functionality via interacting with a particular subunit of PP2A. Mol Cancer Ther; 16(9); 1791–805. ©2017 AACR.

Summary Withania somnifera produces pharmacologically important triterpenoid withanolides that are derived via phytosterol pathway; however, their biosynthesis and regulation remain to be elucidated. A jasmonate- and salicin-inducible WRKY transcription factor from W. somnifera (WsWRKY1) exhibiting correlation with withaferin A accumulation was functionally characterized employing virus-induced gene silencing and overexpression studies combined with transcript and metabolite analyses, and chromatin immunoprecipitation assay. WsWRKY1 silencing resulted in stunted plant growth, reduced transcripts of phytosterol pathway genes with corresponding reduction in phytosterols and withanolides in W. somnifera. Its overexpression elevated the biosynthesis of triterpenoids in W. somnifera (phytosterols and withanolides), as well as tobacco and tomato (phytosterols). Moreover, WsWRKY1 binds to W-box sequences in promoters of W. somnifera genes encoding squalene synthase and squalene epoxidase, indicating its direct regulation of triterpenoid pathway. Furthermore, while WsWRKY1 silencing in W. somnifera compromised the tolerance to bacterial growth, fungal infection, and insect feeding, its overexpression in tobacco led to improved biotic stress tolerance. Together these findings demonstrate that WsWRKY1 has a positive regulatory role on phytosterol and withanolides biosynthesis, and defense against biotic stress, highlighting its importance as a metabolic engineering tool for simultaneous improvement of triterpenoid biosynthesis and plant defense.

In [4], W. Kohnen constructed explicit Shintani lifts from the space of cusp forms of weight $2k$ on $Γ_0(N)$, which are mapped into the plus space $S^+_{k+1/2}(Γ_0(4N))$, where $N$ is an odd integer. This lifting is adjoint to the (modified) Shimura map defined by him with respect to the Petersson scalar product. Using this construction along with the theory of newforms on $S^+_{k+1/2}(Γ_0(4N))$ (where $N$ is an odd square-free natural number) developed in [3], he derived explicit Waldspurger theorem for the newforms belonging to the space $S^+_{k+1/2}(Γ_0(4N))$. Further in this work, Kohnen considered the $\pm 1$ eigen subspaces corresponding to the Atkin-Lehner involution and showed that the intersection of this $\pm$ spaces with the corresponding newform spaces are isomorphic under the Shimura correspondence. A natural question is whether a similar result can be obtained for the intersection of this $(\pm1)$ subspaces with the space of oldforms. In this direction, S. Choi and C. H. Kim [2] considered the case where $N$ is an odd prime $p$ and constructed similar Shimura and Shintani maps between subspaces of forms of half-integral and integral weights. The subspaces considered by Choi and Kim are nothing but $+1$ eigenspace under the Fricke involution. In this paper, we generalise the work of Choi and Kim to the case where $N$ is an odd square-free natural number.

This paper presents a study on replacing the usage of fossil fuel energy with solar energy for lighting the dark and depressing streets of Fugar city, Nigeria. Fugar city is quite populated area without any street lights, almost every house use fossil energy to light up the streets which they access. The main objective is to select best solution among diesel generators, grid electricity, on-site solar photovoltaics and off-site solar photovoltaics. In order to have a sustainable solution for lighting up 210 LED street lights, the four proposed solutions were analyzed based on their technical feasibility, environmental parameters like CO2 emission analysis and cost analysis with simple payback periods. Analysis showed that, on-site solar photovoltaics is best among the other three proposed solutions in terms of technical and financial feasibility with almost negligible emissions leading to sustainability.

India is the TNBC capital of the world due to its highest (31%) prevalence and globally it has huge unmet medical need for its high mortality rate. Aggressive early metastasis is the key for such poor clinical outcome. So, there is a dire need to tackle this deadly disease worldwide by targeting its metastatic process. Epigenetic modulator Enhancer of Zeste homolog 2 (EZH2) promotes hetero-chromatinization of target genes by tri-methylating lysine at 27 of histone H3 (H3K27me3) and it has been shown to be associated with poor prognosis of TNBC. Besides its canonical function as a transcriptional repressor, EZH2 protein has recently been observed to perform H3K27me3 independent functions. Contribution of EZH2 protein (non-canonical) versus H3K27me3 dependent (canonical) function in TNBC pathogenesis remains poorly understood. Here, we aim to dissect the differential role of EZH2 protein versus H3K27me3 in the context of TNBC growth and metastasis. Utilizing stable EZH2 (protein versus hyperactivated H3K27me3) TNBC cells, we discovered that selective hyperactivation of H3K27me3 function of EZH2 but not its protein overexpression markedly promotes cell migration and invasion though cellular proliferation remains unaltered in both the cases. EZH2 protein overexpression (EPOE) and hyperactivation of H3K27me3 (EFHA) in TNBC cells resulted in similar tumor volume and growth pattern in-vivo in an orthotopic TNBC animal model but mice bearing EFHA tumors died significantly earlier than control and EPOE mice. Sequential Luc-GFP/Td tomato based live animal imaging of tumor bearing mice confirmed that EFHA profoundly augments TNBC metastasis compared to control and interestingly, it changes metastatic landscape of TNBC. Control tumors tend to metastasize to lung whereas; EFHA tumors metastasize to spleen first. Splenic metastasis of TNBC shows selective hyperactivation of H3K27me3 as compared to lung and liver metastasis at basal state. EZH2 functional inhibitor EPZ-6438 (currently in Phase-II clinical trial) though did not alter TNBC primary tumor growth but significantly abrogated TNBC metastasis and increased the life expectancy of tumor bearing mice. The high-throughput Illumina RNA-sequencing data identified unique gene signature of EFHA mediated TNBC metastasis corroborating with patient samples and TCGA database. In summary, our data indicate that splenic metastasis of TNBC may be driven by trimethylation function of EZH2 and targeting the methyltransferase activity of EZH2 could be a wonderful strategy to prevent TNBC metastasis in the clinic. Citation Format: Ayushi Verma, Akhilesh Singh, Anup Kumar Singh, Priyank Chaturvedi, Mushtaq Ahmad Nengroo, Krishan kumar Saini, abhipsa Sinha, Dipak Datta. Selective ezh2 functional activation alters the metastatic landscape of triple negative breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3837.

Chemotherapy resistance is one of the pivotal reasons for cancer-associated huge mortality and morbidity. Cancer cells become therapy resistant by smartly evading the execution of apoptosis. Death Receptors (DR) are critical pro-apoptotic factors that regulate cell death, and paucity of death receptors in cancer cells promotes therapy resistance. High expression of CXCR4 is strongly correlated with poor prognosis and adverse clinical outcomes. Inhibitors of the CXCL12-CXCR4 axis, such as FDA approved drug plerixafor (AMD3100), and Nox-A12 have shown limited clinical success in cancer treatment. Alternately, CXCR7, another CXCL12 binding receptor, could be the reason for the limited success of the CXCR4 blockade in clinics. However, recent preclinical data for blocking both the chemokine receptors failed to explain the pro-tumorigenic functions of these axes. Therefore, CXCR4-CXCL12 signaling may not be the exclusive contributor to its pro-tumorigenic functions. In our continuous effort to understand the chemokine receptor signaling inhibition as cancer therapy, here we unexpectedly discovered that instead of CXCR4-CXCL12 signaling, CXCR4 intracellular protein augments therapy resistance and pro-tumorigenic functions. By utilizing stable gain and loss of function approaches, we observed that CXCR4 selectively resists and sensitizes cancer cells against paclitaxel therapy. Our unbiased apoptosis array followed by immunoblot, FACS, real-time PCR, and ChIP analysis clearly demonstrate that CXCR4 promotes downregulation of pro-apoptotic protein DR5 via modulating differential recruitment of transcription factors p53 and YY1 to the promoter of DR5 gene. Inhibiting CXCR4 mediated signals failed to block the above phenotype. Irrespective of CXCR4 surface expression, three different colon cancer xenograft models evidently exhibited loss of CXCR4 protein results in compromised colon tumor growth in vivo. Finally, performing TCGA data mining, real-time PCR, immunofluorescence microscopy of human cancer patient samples and cell lines, we observed that expression of CXCR4 and DR5 are inversely correlated in human cancer. Together, we showed for the first time that targeting intracellular CXCR4 protein may be critical to dampen the pro-tumorigenic functions of CXCR4. Citation Format: Mushtaq Ahmad Nengroo, Shrankhla Maheshwari, Akhilesh Singh, Anup Kumar Singh, Priyank Chaturvedi, Ayushi Verma, Rakesh Kumar Arya, Krishan Kumar Saini, Annapurna Gupta, Anjali Mishra, Dipak Datta. CXCR4 intracellular protein regulates chemotherapy resistance by modulating the expression of death receptor 5 [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6325.

Set7/9 (also known as Set7, Set9, Setd7, and Kmt7) is a lysine methyltransferase that catalyzes the methylation of multiple substrates, including histone H3 and non-histone proteins. Although not essential for normal development and physiology, Set7/9-mediated methylation events play important roles in regulating cellular pathways involved in various human diseases, making Set7/9 a promising therapeutic target. Multiple Set7/9 inhibitors have been developed, which exhibit varying degrees of potency and selectivity in vitro. However, validation of these compounds in vivo has been hampered by the lack of a reliable cellular biomarker for Set7/9 activity. Here, we report the identification of Rpl29, a ribosomal protein abundantly expressed in all cell types, as a major substrate of Set7/9. We show that Rpl29 lysine 5 (Rpl29K5) is methylated exclusively by Set7/9 and can be demethylated by Lsd1 (also known as Kdm1a). Rpl29 is not a core component of the ribosome translational machinery and plays a regulatory role in translation efficiency. Our results indicate that Rpl29 methylation has no effect on global protein synthesis but affects Rpl29 subcellular localization. Using an Rpl29 methylation-specific antibody, we demonstrate that Rpl29K5 methylation is present ubiquitously and validate that (R)-PFI-2, a Set7/9 inhibitor, efficiently reduces Rpl29K5 methylation in cell lines. Thus, Rpl29 methylation can serve as a specific cellular biomarker for measuring Set7/9 activity.

In this paper, we consider certain quaternary quadratic forms and octonary quadratic forms, and by using the theory of modular forms, we find formulae for the number of representations of a positive integer by these quadratic forms.

DNA methylation plays a critical role in the regulation of chromatin structure and gene expression and is involved in a variety of biological processes. The levels and patterns of DNA methylation are regulated by both DNA methyltransferases (DNMT1, DNMT3A and DNMT3B) and ‘demethylating’ proteins, including the ten-eleven translocation (TET) family of dioxygenases (TET1, TET2 and TET3). The effects of DNA methylation on chromatin and gene expression are largely mediated by methylated DNA ‘reader’ proteins, including MeCP2. Numerous mutations in DNMTs, TETs and MeCP2 have been identified in cancer and developmental disorders, highlighting the importance of the DNA methylation machinery in human development and physiology. In this review, we describe these mutations and discuss how they may lead to disease phenotypes.

Curry tree (Murraya koenigii L.) is a rich source of aromatic terpenes and pharmacologically important carbazole alkaloids. Here, M. koenigii leaf transcriptome was generated to gain insight into terpenoid and alkaloid biosynthesis. Analysis of de novo assembled contigs yielded genes for terpene backbone biosynthesis and terpene synthases. Also, gene families possibly involved in carbazole alkaloid formation were identified that included polyketide synthases, prenyltransferases, methyltransferases and cytochrome P450s. Further, two genes encoding terpene synthases (MkTPS1 and MkTPS2) with highest in silico transcript abundance were cloned and functionally characterized to determine their involvement in leaf volatile formation. Subcellular localization using GFP fusions revealed the plastidial and cytosolic localization of MkTPS1 and MkTPS2, respectively. Enzymatic characterization demonstrated the monoterpene synthase activity of recombinant MkTPS1, which produced primarily (−)-sabinene from geranyl diphosphate (GPP). Recombinant MkTPS2 exhibited sesquiterpene synthase activity and formed (E,E)-α-farnesene as the major product from farnesyl diphosphate (FPP). Moreover, mRNA expression and leaf volatile analyses indicated that MkTPS1 accounts for (−)-sabinene emitted by M. koenigii leaves. Overall, the transcriptome data generated in this study will be a great resource and the start point for characterizing genes involved in the biosynthetic pathway of medicinally important carbazole alkaloids.

In this paper, we find the number of representations of the quadratic form $x_1^2+ x_1x_2 + x_2^2 + \ldots + x_{2k-1}^2 + x_{2k-1}x_{2k} + x_{2k}^2,$ for $k=7,9,11,12,14$ using the theory of modular forms. By comparing our formulas with the formulas obtained by G. A. Lomadze, we obtain the Fourier coefficients of certain newforms of level $3$ and weights $7,9,11$ in terms of certain finite sums involving the solutions of similar quadratic forms of lower variables. In the case of $24$ variables, comparison of these formulas gives rise to a new formula for the Ramanujan Tau function., 11 pages, minor revision, certain terms are combined to simplify formulas in Theorem 2.3

Poly ADP-ribose polymerases (PARPs) catalyze massive protein poly ADP-ribosylation (PARylation) within seconds after the induction of DNA single- or double-strand breaks. PARylation occurs at or near the sites of DNA damage and promotes the recruitment of DNA repair factors via their poly ADP-ribose (PAR) binding domains. Several novel PAR-binding domains have been recently identified. Here, we summarize these and other recent findings suggesting that PARylation may be the critical event that mediates the first wave of the DNA damage response. We also discuss the potential for functional crosstalk with other DNA damage-induced post-translational modifications.

In this paper we use the theory of modular forms to find formulas for the number of representations of a positive integer by a certain class of quadratic forms in eight variables, viz., forms of the form \(a_1x_1^2 + a_2 x_2^2 + a_3 x_3^2 + a_4 x_4^2 + b_1(x_5^2+x_5x_6 + x_6^2) + b_2(x_7^2+x_7x_8 + x_8^2)\), where \(a_1\le a_2\le a_3\le a_4\), \(b_1\le b_2\) and \(a_i\)’s \(\in \{1,2,3\}\), \(b_i\)’s \(\in \{1,2,4\}\). We also determine formulas for the number of representations of a positive integer by the quadratic forms \((x_1^2+x_1x_2+x_2^2) + c_1(x_3^2+x_3x_4+x_4^2) + c_2(x_5^2+x_5x_6+x_6^2) + c_3(x_7^2+x_7x_8+x_8^2)\), where \(c_1,c_2,c_3\in \{1,2,4,8\}\), \(c_1\le c_2\le c_3\).

Seminal discoveries have established that epigenetic modifications are important for driving tumor progression. Polycomb group (PcG) proteins are highly conserved epigenetic effectors that maintain, by posttranslational modification of histones, the silenced state of genes involved in critical biologic processes, including cellular development, stem cell plasticity, and tumor progression. PcG proteins are found in two multimeric protein complexes called Polycomb repressive complexes: PRC1 and PRC2. Enhancer of zeste homolog 2 (EZH2), catalytic core subunit of PRC2, epigenetically silences several tumor-suppressor genes by catalyzing the trimethylation of histone H3 at lysine 27, which serves as a docking site for DNA methyltransferases and histone deacetylases. Evidence suggests that overexpression of EZH2 is strongly associated with cancer progression and poor outcome in disparate cancers, including hematologic and epithelial malignancies. The regulatory circuit and molecular cues causing EZH2 deregulation vary in different cancer types. Therefore, this review provides a comprehensive overview on the oncogenic role of EZH2 during tumorigenesis and highlights the multifaceted role of EZH2, as either a transcriptional activator or repressor depending on the cellular context. Additional insight is provided on the recent understanding of the causes and consequences of EZH2 overexpression in specific cancer types. Finally, evidence is discussed on how EZH2 has emerged as a promising target in anticancer therapy and the prospects for targeting EZH2 without affecting global methylation status. Thus, a better understanding of the complex epigenetic regulatory network controlling EZH2 expression and target genes facilitates the design of novel therapeutic interventions. Mol Cancer Res; 12(5); 639–53. ©2014 AACR.

Catharanthus roseus is the sole source of two most important monoterpene indole alkaloid (MIA) anti- cancer agents: vinblastine and vincristine. MIAs possess a terpene and an indole moiety derived from terpenoid and shikimate pathways, respectively. Geranyl diphosphate (GPP), the entry point to the formation of terpene moiety, is a product of the condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) by GPP synthase (GPPS). Here, we report three genes encoding proteins with sequence similarity to large subunit (CrGPPS.LSU) and small subunit (CrGPPS.SSU) of heteromeric GPPSs, and a homomeric GPPSs. CrGPPS.LSU is a bifunctional enzyme producing both GPP and geranyl geranyl diphosphate (GGPP), CrGPPS.SSU is inactive, whereas CrGPPS is a homomeric enzyme forming GPP. Co-expression of both subunits in Escherichia coli resulted in heteromeric enzyme with enhanced activity producing only GPP. While CrGPPS.LSU and CrGPPS showed higher expression in older and younger leaves, respectively, CrGPPS.SSU showed an increasing trend and decreased gradually. Methyl jasmonate (MeJA) treatment of leaves sig- nificantly induced the expression of only CrGPPS.SSU. GFP localization indicated that CrGPPS.SSU is plastidial whereas CrGPPS is mitochondrial. Transient overexpression of AmGPPS.SSU in C. roseus leaves resulted in increased vindoline, immediate monomeric precursor of vinblastine and vincristine. Although C. roseus has both heteromeric and homomeric GPPS enzymes, our results implicate the involvement of only heteromeric GPPS with CrGPPS.SSU regulating the GPP flux for MIA biosynthesis.

Although chemokines are well established to function in immunity and endothelial cell activation and proliferation, a rapidly growing literature suggests that CXC Chemokine receptors CXCR3, CXCR4 and CXCR7 are critical in the development and progression of solid tumors. The effect of these chemokine receptors in tumorigenesis is mediated via interactions with shared ligands I-TAC (CXCL11) and SDF-1 (CXCL12). Over the last decade, CXCR4 has been extensively reported to be overexpressed in most human solid tumors and has earned considerable attention toward elucidating its role in cancer metastasis. To enrich the existing armamentarium of anti-cancerous agents, many inhibitors of CXCL12-CXCR4 axis have emerged as additional or alternative agents for neo-adjuvant treatments and even many of them are in preclinical and clinical stages of their development. However, the discovery of CXCR7 as another receptor for CXCL12 with rather high binding affinity and recent reports about its involvement in cancer progression, has questioned the potential of "selective blockade" of CXCR4 as cancer chemotherapeutics. Interestingly, CXCR7 can also bind another chemokine CXCL11, which is an established ligand for CXCR3. Recent reports have documented that CXCR3 and their ligands are overexpressed in different solid tumors and regulate tumor growth and metastasis. Therefore, it is important to consider the interactions and crosstalk between these three chemokine receptors and their ligand mediated signaling cascades for the development of effective anti-cancer therapies. Emerging evidence also indicates that these receptors are differentially expressed in tumor endothelial cells as well as in cancer stem cells, suggesting their direct role in regulating tumor angiogenesis and metastasis. In this review, we will focus on the signals mediated by this receptor trio via their shared ligands and their role in tumor growth and progression.