32 Background: Triple Negative Breast Cancer (TNBC) accounts for 10-20% of breast cancers and is characterized by poor prognosis, early recurrence, high metastatic potential, and lack of effective targeted therapies. Hypoxia-driven immunosuppression and aberrant tumor vasculature are key hallmarks contributing to TNBC progression and resistance to therapy. The pro-angiogenic chemokine CXCL3, frequently overexpressed in aggressive breast tumors, plays a significant role in abnormal angiogenesis, tumor growth, metastasis, and immune evasion. However, the specific role of tumoral CXCL3 in regulating vascular normalization and immune surveillance remains inadequately understood. This study investigates whether targeting the CXCL3-CXCR2 axis can remodel the tumor microenvironment and enhance immunotherapeutic efficacy in TNBC. Methods: An orthotopic 4T1 murine TNBC model was used to evaluate the impact of CXCL3-CXCR2 axis blockade. Genetic knockdown of CXCL3 and pharmacological inhibition of CXCR2 using SB225002 (5 mg/kg/day, i.p.) were employed. Tumor growth kinetics were monitored. Vascular morphology (CD31, α-SMA), perfusion (lectin), and hypoxia (EF5) were assessed by immunofluorescence. Immune cell profiling was performed using flow cytometry. Proliferation (Ki-67), apoptosis (cleaved caspase-3), phospho-ERK, phospho-p38 signaling, and PD-L1 expression were evaluated. Combination therapy studies were conducted using SB225002 and anti-PD-L1 antibody (200 µg/kg/q3d/i.p.), followed by assessment of tumor progression, lung metastasis, Granzyme B expression, and survival (Kaplan-Meier analysis). Results: CXCR2 inhibition using SB225002 significantly reduced tumor growth and volume (***P < 0.001). Treatment decreased vessel diameter and density while enhancing pericyte coverage, indicating vascular normalization. Improved perfusion and reduced intratumoral hypoxia were observed (*P < 0.05). Importantly, SB225002 enhanced infiltration of anti-tumoral immune cells, including CD8⁺ T cells and M1 macrophages (***P < 0.001), while reducing regulatory T cells (Tregs) and N2 neutrophils. Reduced Ki-67 expression, downregulation of phospho-ERK and phospho-p38 signaling, increased apoptosis, and modulation of PD-L1 expression further supported its anti-tumor efficacy. Combination therapy with SB225002 and anti-PD-L1 demonstrated superior tumor control, reduced lung metastasis, increased Granzyme B expression, and significantly improved survival (*P < 0.05). Conclusions: Targeting the CXCL3-CXCR2 axis in TNBC promotes vascular normalization, reduces hypoxia, enhances anti-tumor immune surveillance, and improves the efficacy of immune checkpoint blockade, offering a promising combinatorial therapeutic strategy.

Chemokines are tiny chemotactic cytokines which play a crucial role in pathophysiology by maintaining homeostasis and inflammation. Their role in the tumour microenvironment is very much puzzling because of both pro- and anti-tumourigenic effects. Chemokines have gained much attention today, since it has been recognized that they are game changers in the TME via controlling immune cell recruitment, angiogenesis, metastasis, tumour growth and drug resistance. In this review, we are exploring the role of several chemokines and their receptors in the TME with special focus on immune cell recruitment, immune surveillance, regulation of immune checkpoints and epithelial mesenchymal transition. We are also reviewing the possibility of targeting chemokines along with immunotherapy for better outcome and disease-free survival. A better understanding on the dual role of chemokine in the TME might help to implement novel therapeutic interventions and adopt precision in targeted therapy.

According to studies, 100% of all artificial implants trigger an immunological response, and 35% of them necessitate a second procedure. The extent to which the chosen biomaterial can navigate biological barriers in vivo determines the success of an implanted biomaterial. The immune system, which is made up of a complex network of cells that trigger an inflammatory response to the biomaterial implanted, is the most important of these barriers. Foreign body response is the collective term for the body’s reaction to implanted material. Provisional matrix development, acute inflammation, and chronic inflammation are some of the conditions that can occur. Injury to the tissue during the implantation procedure, the presence of a foreign body in living tissue, or bacterial infection can all induce these inflammatory reactions. Within minutes of implantation, the implanted material begins to adsorb blood plasma proteins. Monocytes are attracted to these adsorbed proteins, and these monocytes develop into macrophages. The main phagocytic cells recruited by the innate arm of immunity are macrophages. Inflammation can result in the production of granulation tissue, fibrous capsules, and foreign body giant cells, depending on the magnitude of the lesion and the type/role of the biomaterial. Granulation tissue production is the body’s natural attempt to heal an injury through the wound healing process. The recruitment of fibroblasts and myofibroblasts causes a fibrous capsule to grow around the implant, isolating it from the surrounding tissue. Macrophages continue to try to phagocytose the foreign body and produce enzymes and reactive species that worsen the situation. In order to ingest the larger implants, persistent inflammation causes macrophages to polarize and fuse, resulting in multinucleated gigantic cells.

Supplementary Figure 3 - Expression of VEGFR-1 and IL-1ß in TAMs in PAN02 tumors.

Effects of VEGFR-1 signaling ablation on glucose/insulin tolerance and insulin production by the pancreas during obesity.

Supplementary Figure 5 - Effects of VEGFR-1 signaling ablation on PAN02 tumor progression in body-weight matched mice.

Supplementary Figure 1 - A) Vessel density in PAN02 tumors from lean and obese mice, and effect of VEGFR-1-TK-deletion on tumor vessel density and hypoxia markers in obese mice. Supplementary Figure 2 - Gene expression of M1 / M2 markers in TAMs isolated from PAN02 tumors in lean and obese mice.

Breast cancer ranks second among the causes of cancer-related deaths in women. In spite of the recent advances achieved in the diagnosis and treatment of breast cancer, further study is required to overcome the risk of cancer resistance to treatment and thereby improve the prognosis of individuals with advanced-stage breast cancer. The existence of a hypoxic microenvironment is a well-known event in the development of mutagenesis and rapid proliferation of cancer cells. Tumor cells, purposefully cause local hypoxia in order to induce angiogenesis and growth factors that promote tumor growth and metastatic characteristics, while healthy tissue surrounding the tumor suffers damage or mutate. It has been found that these settings with low oxygen levels cause immunosuppression and a lack of immune surveillance by reducing the activation and recruitment of tumor infiltrating leukocytes (TILs). The immune system is further suppressed by hypoxic tumor endothelium through a variety of ways, which creates an immunosuppressive milieu in the tumor microenvironment. Non responsiveness of tumor endothelium to inflammatory signals or endothelial anergy exclude effector T cells from the tumor milieu. Expression of endothelial specific antigens and immunoinhibitory molecules like Programmed death ligand 1,2 (PDL–1, 2) and T cell immunoglobulin and mucin-domain containing-3 (TIM-3) by tumor endothelium adds fuel to the fire by inhibiting T lymphocytes while promoting regulatory T cells. The hypoxic microenvironment in turn recruits Myeloid Derived Suppressor Cells (MDSCs), Tumor Associated Macrophages (TAMs) and T regulatory cells (Treg). The structure and function of newly generated blood vessels within tumors, on the other hand, are aberrant, lacking the specific organization of normal tissue vasculature. Vascular normalisation may work for a variety of tumour types and show to be an advantageous complement to immunotherapy for improving tumour access. By enhancing immune response in the hypoxic tumor microenvironment, via immune-herbal therapeutic and immune-nutraceuticals based approaches that leverage immunological evasion of tumor, will be briefly reviewed in this article. Whether these tactics may be the game changer for emerging immunological switch point to attenuate the breast cancer growth and prevent metastatic cell division, is the key concern of the current study.

Supplementary Table 1: Protein quantification of cytokines in tumors and plasma of WT and Flt1TK-/- obese using ELISA; Supplementary Table 2: Quantification of the expression of IL-1ÃŽÃ,² and VEGFR-1 in TAMs from PAN02 tumors by mmunofluorescence.

Supplementary Tables S1 - S6. Supplementary Table S1. CT values for demosplasia-related genes in PAN02 tumors. Data obtained from PCR array. Supplementary Table S2. CT values for demosplasia-related genes in AK4.4 tumors. Data obtained from PCR array. Supplementary Table S3. Univariate analysis of prognostic factors for resected pancreatic cancer patients with body mass index (BMI) {less than or equal to}25. Supplementary Table S4. Multivariate analysis of prognostic factors for resected pancreatic cancer patients with body mass index (BMI) {less than or equal to}25. Supplementary Table S5. Univariate analysis of prognostic factors for resected pancreatic cancer patients with body mass index (BMI) >25. Supplementary Table S6. Multivariate analysis of prognostic factors for resected pancreatic cancer patients with body mass index (BMI) >25.

Supplementary Figure 7 - Effects of VEGFR-1 signaling ablation on IGF-1R/IR signaling pathways in PAN02 and E0771 tumors implanted in obese mice; Supplementary Figure 8 - Additional effects of VEGFR-1 signaling ablation and metformin on tumor metabolism and vessel density in obese mice.

Supplementary Figure 10 - Additional measurements of PlGF and VEGF-B; Supplementary Figure 11 - Additional effects of PlGF deletion on obesity-induced tumor progression and systemic metabolism.

Effects of VEGFR-1 signaling ablation on E0771 tumor growth, vascular and immune environment in lean and obese mice.

Effects of VEGFR-1 signaling ablation on body weight gain, immune cell infiltration and vasculature in adipose tissues during obesity.

Introduction: Isodeoxyelephantopin (C19H20O6), plant-derived sesquiterpene, extracted from Elephantopus Scaber Linn., have tremendous contributions in inhibiting the proliferation and metastasis of cancer cells. Materials and Methods: The cell viability assay was shown to be procured half-maximal inhibitory concentration of isodeoxyelephantopin on the MDA-MB-231cell line. Apoptosis was screened by AO/ EB staining and it was confirmed by annexin V staining. The DNA damaging property of IDOE was examined by comet assay. The cell cycle arrest determined using flow cytometry analysis and the growth inhibition due to the modulation of cell cycle regulatory protein p53 were substantiated by RT PCR. Scratch wound migratory assay was done to evaluate the migratory effect of IDOE on the MDA-MB-231 cell line. RT-PCR analysis showed that expression of MMP-2/9 was inhibited by IDOE treatment. Result: Anti-proliferative and anti-migratory effect of IDOE was determined by cell cycle analysis and annexin v staining. Cells were arrested in the G2/M phase and it is due to the modulation of cell cycle regulatory protein p53. The mRNA level expression studies of matrix metalloproteinases, MMP-2/9 provide supporting data to prove the anti-migratory property.

Anti-vascular endothelial growth factor (VEGF) therapy has failed to improve survival in patients with breast cancer (BC). Potential mechanisms of resistance to anti-VEGF therapy include the up-regulation of alternative angiogenic and proinflammatory factors. Obesity is associated with hypoxic adipose tissues, including those in the breast, resulting in increased production of some of the aforementioned factors. Hence, we hypothesized that obesity could contribute to anti-VEGF therapy's lack of efficacy. We found that BC patients with obesity harbored increased systemic concentrations of interleukin-6 (IL-6) and/or fibroblast growth factor 2 (FGF-2), and their tumor vasculature was less sensitive to anti-VEGF treatment. Mouse models revealed that obesity impairs the effects of anti-VEGF on angiogenesis, tumor growth, and metastasis. In one murine BC model, obesity was associated with increased IL-6 production from adipocytes and myeloid cells within tumors. IL-6 blockade abrogated the obesity-induced resistance to anti-VEGF therapy in primary and metastatic sites by directly affecting tumor cell proliferation, normalizing tumor vasculature, alleviating hypoxia, and reducing immunosuppression. Similarly, in a second mouse model, where obesity was associated with increased FGF-2, normalization of FGF-2 expression by metformin or specific FGF receptor inhibition decreased vessel density and restored tumor sensitivity to anti-VEGF therapy in obese mice. Collectively, our data indicate that obesity fuels BC resistance to anti-VEGF therapy via the production of inflammatory and angiogenic factors.

Abstract It remains unclear how obesity worsens treatment outcomes in patients with pancreatic ductal adenocarcinoma (PDAC). In normal pancreas, obesity promotes inflammation and fibrosis. We found in mouse models of PDAC that obesity also promotes desmoplasia associated with accelerated tumor growth and impaired delivery/efficacy of chemotherapeutics through reduced perfusion. Genetic and pharmacologic inhibition of angiotensin-II type-1 receptor reverses obesity-augmented desmoplasia and tumor growth and improves response to chemotherapy. Augmented activation of pancreatic stellate cells (PSC) in obesity is induced by tumor-associated neutrophils (TAN) recruited by adipocyte-secreted IL1β. PSCs further secrete IL1β, and inactivation of PSCs reduces IL1β expression and TAN recruitment. Furthermore, depletion of TANs, IL1β inhibition, or inactivation of PSCs prevents obesity-accelerated tumor growth. In patients with pancreatic cancer, we confirmed that obesity is associated with increased desmoplasia and reduced response to chemotherapy. We conclude that cross-talk between adipocytes, TANs, and PSCs exacerbates desmoplasia and promotes tumor progression in obesity. Significance: Considering the current obesity pandemic, unraveling the mechanisms underlying obesity-induced cancer progression is an urgent need. We found that the aggravation of desmoplasia is a key mechanism of obesity-promoted PDAC progression. Importantly, we discovered that clinically available antifibrotic/inflammatory agents can improve the treatment response of PDAC in obese hosts. Cancer Discov; 6(8); 852–69. ©2016 AACR. See related commentary by Bronte and Tortora, p. 821. This article is highlighted in the In This Issue feature, p. 803

Abstract Purpose: Obesity promotes pancreatic and breast cancer progression via mechanisms that are poorly understood. Although obesity is associated with increased systemic levels of placental growth factor (PlGF), the role of PlGF in obesity-induced tumor progression is not known. PlGF and its receptor VEGFR-1 have been shown to modulate tumor angiogenesis and promote tumor-associated macrophage (TAM) recruitment and activity. Here, we hypothesized that increased activity of PlGF/VEGFR-1 signaling mediates obesity-induced tumor progression by augmenting tumor angiogenesis and TAM recruitment/activity. Experimental Design: We established diet-induced obese mouse models of wild-type C57BL/6, VEGFR-1 tyrosine kinase (TK)-null, or PlGF-null mice, and evaluated the role of PlGF/VEGFR-1 signaling in pancreatic and breast cancer mouse models and in human samples. Results: We found that obesity increased TAM infiltration, tumor growth, and metastasis in pancreatic cancers, without affecting vessel density. Ablation of VEGFR-1 signaling prevented obesity-induced tumor progression and shifted the tumor immune environment toward an antitumor phenotype. Similar findings were observed in a breast cancer model. Obesity was associated with increased systemic PlGF, but not VEGF-A or VEGF-B, in pancreatic and breast cancer patients and in various mouse models of these cancers. Ablation of PlGF phenocopied the effects of VEGFR-1-TK deletion on tumors in obese mice. PlGF/VEGFR-1-TK deletion prevented weight gain in mice fed a high-fat diet, but exacerbated hyperinsulinemia. Addition of metformin not only normalized insulin levels but also enhanced antitumor immunity. Conclusions: Targeting PlGF/VEGFR-1 signaling reprograms the tumor immune microenvironment and inhibits obesity-induced acceleration of tumor progression. Clin Cancer Res; 22(12); 2993–3004. ©2016 AACR.

Arginine, one among the 20 most common natural amino acids, has a pivotal role in cellular physiology as it is being involved in numerous cellular metabolic and signaling pathways. Dependence on arginine is diverse for both tumor and normal cells. Because of decreased expression of argininosuccinate synthetase and/or ornithine transcarbamoylase, several types of tumor are auxotrophic for arginine. Deprivation of arginine exploits a significant vulnerability of these tumor cells and leads to their rapid demise. Hence, enzyme-mediated arginine depletion is a potential strategy for the selective destruction of tumor cells. Arginase, arginine deiminase and arginine decarboxylase are potential enzymes that may be used for arginine deprivation therapy. These arginine catabolizing enzymes not only reduce tumor growth but also make them susceptible to concomitantly administered anti-cancer therapeutics. Most of these enzymes are currently under clinical investigations and if successful will potentially be advanced as anti-cancer modalities.