Editorial: Metabolic Regulation Under Oxidative Stress in Cancer

Oxidative stress has emerged as a key component of cancer metabolism that impacts multiple facets of tumor biology (1). Recent studies have shed light on the complex interplay of cellular redox and its impact on molecular mechanisms that govern metabolic reprogramming under oxidative stress (2, 3). It is well established that altered glucose metabolism exhibited by tumor cells leads to an enormous oxidative burden through various metabolic routes (4, 5). What is less known but remains a great interest to the field is whether elevated oxidative stress has a causal role in the development of aggressive and resistant tumor phenotypes (6). To that end, this Research Topic explores various metabolic routes that protect tumor cells against oxidative stress in diverse cancer models. It is anticipated that the development of advanced therapeutic approaches requires an in-depth understanding of the oxidative stress-mediated metabolic processes in cancer, including its effects on aspects of the tumor milieu such as immune cells-related functions

IL10 variant g.5311A is associated with Visceral Leishmaniasis in Indian population

Background: Visceral Leishmaniasis (VL) is a multifactorial disease, where the host genetics play a significant role in determining the disease outcome. The immunological role of anti-inflammatory cytokine, Interleukin 10 (IL10), has been well-documented in parasite infections and considered as a key regulatory cytokine for VL. Although VL patients in India display high level of IL10 in blood serum, no genetic study has been conducted to assess the VL susceptibility/resistance. Therefore, the aim of this study is to investigate the role of IL10 variations in Indian VL; and to estimate the distribution of disease associated allele in diverse Indian populations. Methodology: All the exons and exon-intron boundaries of IL10 were sequenced in 184 VL patients along with 172 ethnically matched controls from VL endemic region of India. Result and Discussion: Our analysis revealed four variations; rs1518111 (2195 A>G, intron), rs1554286 (2607 C>T, intron), rs3024496 (4976 T>C, 3’ UTR) and rs3024498 (5311 A>G, 3’ UTR). Of these, a variant g.5311A is significantly associated with VL (χ2 = 18.87; p = 0.00001). In silico approaches have shown that a putative micro RNA binding site (miR-4321) is lost in rs3024498 mRNA. Further, analysis of the above four variations in 1138 individuals from 34 ethnic populations, representing different social and linguistic groups who are inhabited in different geographical regions of India, showed variable frequency. Interestingly, we have found, majority of the tribal populations have low frequency of VL (‘A’ of rs3024498); and high frequency of leprosy (‘T’ of rs1554286), and Behcet’s (‘A’ of rs1518111) associated alleles, whereas these were vice versa in castes. Our findings suggest that majority of tribal populations of India carry the protected/less severe allele against VL, while risk/more severe allele for leprosy and Behcet’s disease. This study has potential implications in counseling and management of VL and other infectious diseases

TNIK Regulation of Interferon Signaling and Endothelial Cell Response to Virus Infection

BACKGROUND: Traf2 and Nck-interacting kinase (TNIK) is known for its regulatory role in various processes within cancer cells. However, its role within endothelial cells (ECs) has remained relatively unexplored. METHODS: Leveraging RNA-seq data and Ingenuity Pathway Analysis (IPA), we probed the potential impact of TNIK depletion on ECs. RESULTS: Examination of RNA-seq data uncovered more than 450 Differentially Expressed Genes (DEGs) in TNIK-depleted ECs, displaying a fold change exceeding 2 with a false discovery rate (FDR) below 0.05. IPA analysis unveiled that TNIK depletion leads to the inhibition of the interferon (IFN) pathway [-log ( SUMMARY: Our findings suggest that TNIK plays a crucial role in regulating the EC response to virus infections through modulation of the IFN pathway

Supplemental Oxygen Protects Heart Against Acute Myocardial Infarction

Myocardial infarction (MI), which occurs often due to acute ischemia followed by reflow, is associated with irreversible loss (death) of cardiomyocytes. If left untreated, MI will lead to progressive loss of viable cardiomyocytes, deterioration of cardiac function, and congestive heart failure. While supplemental oxygen therapy has long been in practice to treat acute MI, there has not been a clear scientific basis for the observed beneficial effects. Further, there is no rationale for the amount or duration of administration of supplemental oxygenation for effective therapy. The goal of the present study was to determine an optimum oxygenation protocol that can be clinically applicable for treating acute MI. Using EPR oximetry, we studied the effect of exposure to supplemental oxygen cycling (OxCy) administered by inhalation of 21–100% oxygen for brief periods (15–90 min), daily for 5 days, using a rat model of acute MI. Myocardial oxygen tension (pO2), cardiac function and pro-survival/apoptotic signaling molecules were used as markers of treatment outcome. OxCy resulted in a significant reduction of infarct size and improvement of cardiac function. An optimal condition of 30-min OxCy with 95% oxygen + 5% CO2 under normobaric conditions was found to be effective for cardioprotection

Role of TRAF2 and NCK Interacting Kinase (TNIK) in the Regulation of Innate Interferon Signaling and Suppression of Viral Infection in Endothelial Cells

Endothelial cells (ECs) are the major cell types that govern vascular tone and homeostasis. Endothelial inflammation and activation are the first step in the initiation of atherosclerosis and several vascular dysfunctions. Interferon signaling (IFN Type I and II) are key inflammatory pathways in ECs and are implicated to be involved in the vascular complications associated with viral infections and systemic lupus erythematosus. Traf2 and Nck-interacting kinase (TNIK), is a serine/threonine-protein kinase (Ste20) family member of MAP kinase kinase kinase kinases (MAP4K). TNIK is reported to be involved in multiple biological functions such as regulation of actin dynamics, cell morphology, neuronal structure, and cellular proliferation. The underlying mechanisms of TNIK in EC functions and inflammation remains largely unknown. To identify the pathways and functions of TNIK in ECs, we performed next-generation RNA sequencing (RNA-seq) in ECs under TNIK depletion conditions (siTNIK). We found that TNIK affects several cellular pathways, but most significantly they were involved in the regulation of interferon (IFN) and hypercytokinemia/hyperchemokinemia pathways. Using molecular expression studies, we identified that TNIK depletion significantly downregulates several interferon molecules at both mRNA and protein expression levels. Further, to probe the regulatory effects induced by TNIK on the IFN pathway we used treatment of wildtype human adenovirus to ECs. Adenovirus have been previously shown to elicit strong interferon responses in many human cell types. We observed cytopathic effect (CPE) induced by wild type virus only under siTNIK condition but not in other controls. In addition, after the treatment of wildtype adenovirus, viral replication was observed only under TNIK depletion but not in other controls, suggesting that TNIK plays an important role in activation of IFN pathway and in the suppression of viral infection in ECs. In addition to elucidating the novel role of TNIK in activation of interferon pathway, our study supports the idea that ECs have innate immune functions manifested by strong interferon responses as a pro-survival mechanism. Our study will pave the way for the understanding of chronic vascular conditions under several inflammatory conditions and viral infections and be critical in the development of therapeutics targeting vascular dysfunctions

436 Immunotherapy Sensitization via Tumor Acidosis Mitigation by Esomeprazole Monitored with MRI

OBJECTIVES/GOALS: Acidity and the lactate-to-pyruvate ratio correlate with immunotherapy resistance. AcidoCEST MRI and hyperpolarized magnetic resonance spectroscopy (HP-MRS) measure extracellular pH and lactate-to-pyruvate ratio. We will establish a baseline for these biomarkers then observe changes after combination esomeprazole and immunotherapy. METHODS/STUDY POPULATION: We used multiple melanoma models created via serial in vivo passage under immunotherapeutic pressure (FVAX, CTLA-4, PD-1, PD-L1). We used four of these corresponding to 25%, 50%, 75% and 100% resistance (TMT, F2, F3, and F4, respectively). HP-MRS was performed two weeks post implantation in male BL6 mice with AcidoCEST MRI 2-3 days later. Tumors were implanted in additional mice and grown for 1 week. We used esomeprazole as a possible immunotherapy sensitizer. Esomeprazole (or PBS) alone and in combination with immune checkpoint blockade (ICB; αCTLA-4, αPD-1) was then conducted every 3 days for 3 doses. ICB was administered 3h after esomeprazole. AcidoCEST MRI was performed the day after the final dose of combination therapy and 3h after esomeprazole (or PBS) alone. HP-MRS was performed 2-3 days after acidoCEST MRI. RESULTS/ANTICIPATED RESULTS: There was a statistical increase in the lactate-to-pyruvate ratio of the F4 group compared with TMT, F2, and F3 groups (p < 0.05). The TMT, F2, and F3 groups did not differ significantly. The extracellular pH (pHe) of the TMT group was statistically lower than the F2 and F4 groups (p < 0.05). The pHe did not differ significantly between the TMT and F3 groups nor the F2, F3, and F4 groups. The lactate-to-pyruvate ratio and pHe after combination treatment with esomeprazole and ICB did not differ compared to PBS+ICB control. Treatment with esomeprazole alone generated higher lactate-to-pyruvate ratio compared with PBS alone. Tumor volume curves and survival curves of mice bearing F4 tumors treated with esomeprazole combination with ICB showed no difference compared with PBS+ICB, PBS alone, and esomeprazole alone. DISCUSSION/SIGNIFICANCE: We differentiated between the 100% and 25% resistant models with both pHe and lactate-to-pyruvate ratio, although the pHe was counterintuitive. Esomeprazole was ineffective, but other potential sensitizers exist. A non-invasive clinical imaging tool and sensitizer would permit more personalized treatment plans so treatment is more effective

Disrupted stiffness ratio alters nuclear mechanosensing

International audienc

The structure of the human <i>IL10</i> (chr1, 206945839–206940947; ENST00000423557).

<p>Exons of the gene are shown in pink, introns in brown. rs1518111 (2195 A>G) and rs1554286 (2607 C>T) were the intronic variant of second and third exons while rs3024496 (4976 T>C) and rs3024498 (5311 A>G) were the 3’ UTR variant of fifth exons.</p

A. Distribution of genotype frequencies for<i>IL10</i> polymorphism, rs1518111 A>G; rs1554286 C>T; rs3024496 C>T and rs3024498 A>G in VL case and control subjects

<p>. B. Distribution of allele frequencies for <i>IL10</i> polymorphism, rs1518111 A>G; rs1554286 C>T; rs3024496 C>T and rs3024498 A>G in VL case and control subjects.</p