Angiogenesis Signaling Pathways Research Articles

SPDR-5 ESTABLISHMENT OF A CONTINUOUS CULTURE METHOD FOR MENINGIOMAS AND THE EFFECTS OF THE GREM2-BMP SIGNALLING PATHWAY ON PROLIFERATION AND ANGIOGENESIS

Abstract Purpose Meningiomas are closely related to osteogenesis, including calcification and hyperostosis at the site of attachment. Signaling pathways involved in tumor formation and extension of meningioma have not yet been elucidated. Treatment options include surgical removal and radiotherapy, but there are no reports of effective drug treatment. This is due to the difficulty of culturing non-malignant meningiomas, which has made basic experiments difficult. In this study, we established a continuous culture method for meningiomas and investigated the signaling pathways involved in meningioma growth. Methods 67 consecutive cases of meningiomas treated in our department for three years from Apr, 2019 to March, 2022. Primary continuous culture was performed by modifying the conditional reprogramming (CR) method. Sixty-four cases were studied, excluding three cases in which culture was abandoned due to contamination. The mean age was 62.6 years, Grade 3 (6%), Grade 2 (14%) and Grade 1 (80%). These were used to observe morphology, proliferation and genetic analysis involved in angiogenesis. Results 1) Sustained culture lasted an average of 150 days with no differences between Grades; 2) Inhibition of BMP signaling involved in osteogenesis suppressed cell proliferation regardless of Grade, and the IC50 of the inhibitor suggested the involvement of the receptor ALK3; 3) We identified GREM2 as upstream regulator of BMP signals 4) GREM2 is highly expressed in menigotherial meningiomas and immunostaining showed strong expression around whole and calcified areas. 5) GREM2 expression is negatively correlated with MIB-1 L.I., so overexpression of GREM2 in malignant meningioma cells decreased ID, which is involved in proliferation downstream of BMP signaling, and increased angiogenic factor VEGFA, which is involved in proliferation. Conclusion A method for the continuous culture of meningiomas was established. Using this experimental system, we identified the involvement of the GREM2-BMP signaling pathway in proliferation and angiogenesis in meningiomas as well as calcification.

Nanotherapy: New Approach for Impeding Hepatic Cancer Microenvironment via Targeting Multiple Molecular Pathways.

Hepatocellular carcinoma (HCC) microenvironment has been recognized as a key contributor for cancer progression, metastasis, and drug resistance. The crosstalk between tumor cells, the vascular endothelial growth factor (VEGF), and the chemokine (C-C motif) ligand 2 (CCL2) signaling networks mediates immunoinhibitory impact and facilitates tumor angiogenesis. The current investigation aimed at exploring the potent anti-cancer activity of the newly designed nano-based anti-cancer therapy comprising anti-VEGF drug, avastin (AV), and CCR2 antagonist (CR) to counteract HCC and tracking its mode of action in vivo. The prepared AV, CR, and AVCR nanoprototypes were characterized by nanoscale characterization techniques in our previous work. Here, they are applied for unearthing their anti-cancer properties / mechanisms in hepatic cancer-induced rats via analyzing protein levels and genetic expression of the elements incorporated in the angiogenesis, apoptosis, and metastasis signalling pathways. The present results revealed a significant down-regulation in the angiogenesis, survival and metastasis indices along with up-regulation in the pro-apoptotic mediators upon treatment of hepatic cancer-bearing rats with the novel synthesized nanomaterials when compared with the untreated counterparts. We showed across HCC model that anti-VEGF in combination with CCR2 antagonism therapy leads to sensitization and enhanced tumor response over anti-VEGF or CCR2 antagonism monotherapy, particularly in its nanoscale formulation. The present approach provides new mechanistic insights into the powerful anti-hepatic cancer advantage of the novel nanoprototypes which is correlated with modulating critical signal transduction pathways implicated in tumor microenviroment such as angiogenesis, apoptosis and metastasis. This research work presents a substantial foundation for future studies focused on prohibiting cancer progression and recovery by targeting tumor microenviroment.

Mohawk impedes angiofibrosis by preventing the differentiation of tendon stem/progenitor cells into myofibroblasts

Adult tendons heal via fibrovascular scarring with inferior biomechanical properties. Mohawk (Mkx) emerged as a pivotal actor in tenolineage commitment. However, its precise function in tendinopathy remains poorly understood. This study investigates the cellular and molecular mechanisms underlying Mkx’ role in fibrovascular healing. Human samples were collected to test fibrovascular markers. We then performed RNAseq on Mkx−/− mice compared to their wild type littermates to decipher Mkx regulome. We therefore sought to reproduce TSPCs transition to myofibroblasts in-vitro by over-expressing MyoD and followed by phenotypic and experimental cells’ characterization using microscopy, qRT-PCR, flow cytometry sorting, presto-blue cell viability assay and immunofluorescence. Two different in vivo models were used to assess the effect of the MyoD-expressing myofibroblasts: transplantation in the dorsal area of immunodeficient mice and in an adult Achilles tendon injury model. To prevent angiofibrosis, we tested the molecule Xav939 and proceeded with histological stainings, q-RT PCR transcriptional quantification of angifibrotic markers, mechanical tests, and immunofluorescence. Tendinopathy samples showed fibrovascular healing with decreased tenolineage phenotype. Transcriptomic analysis of Mkx−/− tendons revealed myofibroblast-associated biological processes. Over-expression of MyoD in WT tendon stem progenitor cells (TSPCs) gave rise to myofibroblasts reprogramming in-vitro and fibrovascular scarring in-vivo. MKX directly binds to MyoD promoter and underlies global regulative processes related to angiogenesis and Wnt signaling pathway. Blocking Wnt signaling with the small molecule Xav393 resulted in higher histological and biomechanical properties. Taken together, our data provide the first in vivo and in-vitro evidence of tendon stem progenitor cells to myofibroblasts transition and show improved tendon healing via angiofibrosis modulation, thus opening potential therapeutic avenues to treat tendinopathy patients.

A role for steroid 5 alpha-reductase 1 in vascular remodeling during endometrial decidualization.

Decidualization is the hormone-dependent process of endometrial remodeling that is essential for fertility and reproductive health. It is characterized by dynamic changes in the endometrial stromal compartment including differentiation of fibroblasts, immune cell trafficking and vascular remodeling. Deficits in decidualization are implicated in disorders of pregnancy such as implantation failure, intra-uterine growth restriction, and pre-eclampsia. Androgens are key regulators of decidualization that promote optimal differentiation of stromal fibroblasts and activation of downstream signaling pathways required for endometrial remodeling. We have shown that androgen biosynthesis, via 5α-reductase-dependent production of dihydrotestosterone, is required for optimal decidualization of human stromal fibroblasts in vitro, but whether this is required for decidualization in vivo has not been tested. In the current study we used steroid 5α-reductase type 1 (SRD5A1) deficient mice (Srd5a1-/- mice) and a validated model of induced decidualization to investigate the role of SRD5A1 and intracrine androgen signaling in endometrial decidualization. We measured decidualization response (weight/proportion), transcriptomic changes, and morphological and functional parameters of vascular development. These investigations revealed a striking effect of 5α-reductase deficiency on the decidualization response. Furthermore, vessel permeability and transcriptional regulation of angiogenesis signaling pathways, particularly those that involved vascular endothelial growth factor (VEGF), were disrupted in the absence of 5α-reductase. In Srd5a1-/- mice, injection of dihydrotestosterone co-incident with decidualization restored decidualization responses, vessel permeability, and expression of angiogenesis genes to wild type levels. Androgen availability declines with age which may contribute to age-related risk of pregnancy disorders. These findings show that intracrine androgen signaling is required for optimal decidualization in vivo and confirm a major role for androgens in the development of the vasculature during decidualization through regulation of the VEGF pathway. These findings highlight new opportunities for improving age-related deficits in fertility and pregnancy health by targeting androgen-dependent signaling in the endometrium.

Endothelium dysfunction in hind limb arteries of male Zucker Diabetic-Sprague Dawley rats

Endothelium dysfunction produces peripheral vascular disease comorbidities in type 2 diabetes, including hypertension, and critical limb ischemia. In this study we aimed to test endothelial dysfunction, the vasodilator effects of a proteinase-activated receptor 2 (PAR2) agonist (2fLIGRLO), and thromboxane A2 synthase inhibitor (ozagrel) on PAR2 vasodilation in hind limb arteries ex vivo, using Zucker Diabetic-Sprague Dawley (ZDSD) rats, a model of type 2 diabetes. Male Sprague Dawley rats (SD) and ZDSD were fed a high-fat content ‘Western diet’ from 16 to 20 weeks of age (wks) then fed a standard laboratory diet. We identified diabetic ZDSD rats by two consecutive blood glucose measurements > 12.5 mM, based on weekly monitoring. We used acetylcholine, 2fLIGRLO, and nitroprusside with wire-myograph methods to compare relaxations of femoral, and saphenous arteries from diabetic ZDSD (21–23 wks) to age-matched normoglycemic SD. All arteries showed evidence of endothelium dysfunction using acetylcholine (reduced maximum relaxations, reduced sensitivity), and higher sensitivities to 2fLIGRLO, and nitroprusside in ZDSD vs SD. Ozagrel treatment of ZDSD distal segments, and end-branches of saphenous arteries decreased their sensitivities to 2fLIGRLO. We tested aortas for altered expression of endothelium-specific gene targets using PCR array and qPCR. PAR2, and placental growth factor gene transcripts were 1.5, and 4-times higher in ZDSD than SD aortas. Hind limb arteries of ZDSD exhibit endothelium dysfunction having less GPCR agonist induced vasodilation by endothelial NO-release. Different expression of several endothelial genes in ZDSD vs SD aortas, including PAR2, suggests altered inflammatory, and angiogenesis signaling pathways in the endothelium of ZDSD.

SLC7A5 is a lung adenocarcinoma-specific prognostic biomarker and participates in forming immunosuppressive tumor microenvironment

BackgroundAmino acid metabolism participates in forming immunosuppressive tumor microenvironment. Amino acid transporters (AATs), as a gate for admission, remains to be studied. Materials and methodsWe identified LUAD-specific prognostic AATs, SLC7A5 by differential expression analysis, logistic regression, machine learning, Kaplan-Meier analysis, AUC value filtrating and Cox regression. Then differential expression and distribution of SLC7A5 were depicted. Copy number variation, DNA methylation, transcriptional factors and ceRNA network were investigated to explore potential mechanism causing differential expression. The prognostic and clinical relation were evaluated by Kaplan-Meier analysis, Cox regression analysis. GSEA and GSVA were used to analyze altered pathways between SLC7A5 high- and low-groups. The expression of HLA-related genes and immune checkpoint genes, and immune cells infiltration were detected. SLC7A5 expression in immune cells was evaluated by single-cell sequencing data. IPS and an independent immunotherapy cohort assessed response rates of patients with distinct SLC7A5 expression. Proliferation assay and wound healing assay validated the effects of SLC7A5 on proliferation and migration of LUAD cells. Western blotting and cell viability assays were performed to detect mTORC1 pathway activity and sensitivity to rapamycin. ResultsSLC7A5 was a LUAD-specific prognostic AAT and had significant differential expression in transcription and translation level. Methylation levels of cg00728300, cg00858400, cg12408911, cg08710629 were negative correlation with SLC7A5 expression. FOXP3 and TFAP2A were possible transcription factors and miR-30a-5p, miR-184, miR-195-5p may target SLC7A5 mRNA. SLC7A5 high-expression indicated poor prognosis and was an independent prognostic factor. mTORC1, cell cycle, DNA damage repair, response to reactive oxygen, angiogenesis, epithelial-mesenchymal transition (EMT) and various growth factors signaling pathways were activated in SLC7A5 high-expression group. Interestingly, SLC7A5 high-expression group had less immune-related genes expression and immune cells infiltration. Single-cell sequencing data also suggested SLC7A5 was downregulated in various T cells, especially effector T cells. Moreover, high SLC7A5 expression indicated poor immunotherapy efficacy and higher sensitivity to inhibitors of mTORC1 pathway, cell cycle and angiogenesis. SLC7A5 deficiency abrogated proliferation, migration and mTORC1 pathway activity. ConclusionsIn summary, as a LUAD-specific prognostic AAT, SLC7A5 is involved in activation of multiple oncogenic pathways and indicates poor prognosis. Moreover, SLC7A5 may participate in forming immunosuppressive TME and is associated with low response of immunotherapy. SLC7A5 is promising to be a new diagnostic and prognostic biomarker and therapeutic target in LUAD.

Identification of immune subtypes to guide immunotherapy and targeted therapy in clear cell renal cell carcinoma.

Accumulating pieces of evidence suggested that immunotypes may indicate the overall immune landscape in the tumor microenvironment, which were closely related to therapeutic response. The purpose of this study was to classify and define the immune subtypes of clear cell renal cell carcinoma (ccRCC), so as to authenticate the potential immune subtypes that respond to immunotherapy. Transcriptome expression profile and mutation profile data of ccRCC, as well as clinical characteristics used in this study were obtained from TCGA database. There were significant differences in the infiltration of immune cells, immune checkpoints, and antigens between ccRCC and para-cancerous tissues. According to immune components, patients with ccRCC were divided into three immune subtypes, with different clinical and molecular characteristics. Compared with other subtypes, IS2 showed cold immune phenotype, and was associated with better survival. IS1 represented complex immune populations and was associated with poor overall survival (OS) and progression free survival (PFS). Further analysis indicated that expression of immune checkpoints also differed among the three subtypes, and was abnormally up-regulated in IS3. Pathway enrichment analysis indicated that the mTOR signaling pathway was abnormally enriched in IS3, while the TGF_BETA, ANGIOGENESIS and receptor tyrosine kinase signaling pathways were abnormally enriched in IS2. Furthermore, there was an abnormal enrichment of the epithelial-to-mesenchymal transition (EMT) signaling pathway in IS1, which may be associated with a higher rate of metastasis. Finally, SCG2 was screened as a specific antigen of ccRCC, which was not only related to poor prognosis, but also significantly associated with immune cells and immune checkpoints. In conclusion, the immune subtypes of ccRCC may provide new insights into the tumor biology and the precise clinical management of this disease.

The regulatory role of HOX interacting lncRNA in oral cancer-An in silico analysis.

We aim to elucidate the interaction of long noncoding RNAs with HOX genes and their regulatory role and potential drug candidates in oral cancer. The interaction network was constructed using RNA Interactome and the RNA Interactome from the Sequencing Experiments database. The differential expression of HOX genes and HOX interacting lncRNAs was assessed using the TCGA-Head and Neck Squamous Cell Carcinoma oral cancer dataset using DESeq2 R-package. Further, the functional enrichment analysis was performed for the differentially expressed HOX genes and HOX-interacting lncRNAs using Gene Ontology, long noncoding RNA Set Enrichment Analysis, lncRNA ontology annotation extractor and repository (Lantern), and LncRNA Ontology tools. Drug-lncRNA interaction and the effect of drugs on lncRNA expression were assessed from the D-lnc tool. A total of 78 unique interactions were identified between HOX and lncRNAs. Differential expression analysis showed 27 HOX genes and 10 HOX-interacting lncRNAs in oral cancer. HOX genes and HOX-interacting lncRNAs were involved in crucial regulatory processes like cell cycle regulation, cell proliferation and migration, epithelial-mesenchymal transition, angiogenesis, and cell signaling pathways. Cancer hallmark analysis from using long noncoding RNA Set Enrichment Analysis showed the involvement of HOTAIR, HOTTIP MIR503HG, and CDKN2B-AS1 in proliferation, migration, and invasion. Panobinostat was the common drug that influenced the expression of HOTAIR, HOTAIRM1, HOTTIP and CDKN2B-AS1. Differentially expressed HOX-interacting lncRNAs are involved in various regulatory biological processes and cancer hallmark events in oral cancer. The creation of interaction networks may expand the existing knowledge of oral cancer signaling pathways and the discovery of novel targets.

Biological response of extracorporeal shock wave therapy to tendinopathy in vivo (review).

Tendinopathy is a degenerative disease of the tendons caused by prolonged overstretching or overuse of the tendons. It accounts for a large proportion of musculoskeletal disorders which can occur in all age groups. The management of tendinopathy is typically conservative. In clinical practice, when other conservative treatments fail, extracorporeal shock wave therapy (ESWT) is normally used as an efficient alternative to surgical management. Several basic studies have shown that ESWT with lower energy flux densities can produce some biological responses in vivo to tendinopathy and may accelerate the initiation of the healing process in injured tendons. ESWT has a positive impact on the interactive chain of biological response, enhancing the signaling pathways of angiogenesis through mechanical conduction, and promoting cell proliferation and collagen formation. Finally, it helps tissue regeneration by controlling inflammation. The purpose of this review is to summarize the biological responses generated by ESWT in tendinopathy through a comprehensive review of the published literature. Although ESWT has been used clinically for the treatment of tendinopathies for nearly decades, less is known about the experimental studies of its biological effects on tendon tissue. Further studies on the biological response of ESWT for tendon injuries in vivo are needed in the future in order to provide better management to patients.

Itrifal-e-Aftimoon potentiates imatinib-induced anti-leukemic effect by influencing FAK/STAT/Akt/ERK signalling pathways against chronic myeloid leukaemia in vitro.

Limited treatment options are available for advanced stages of chronic myeloid leukaemia (CML). Moreover, patients' relapse after a short remission period, which prompts them to identify a potent drug with the least toxicity. An Unani herbal formulation, Itrifal-e-Aftimoon (IEA) is used for certain neurological disorders, however, its antitumor potential has not been reported yet in any malignancy, including CML. The aqueous extract of IEA was characterized by HPLC/LC-MS and used alone or in combination with standard drug, imatinib in CML cell lines (K562, KU812) in vitro to assess its effect on cancer-associated parameters such as cytotoxicity, cell cycle, apoptosis, oxidative stress, inflammation, angiogenesis, and certain signalling pathways. LC-MS characterization of IEA showed the presence of antitumor compounds including catechin and caffeic acid. Treatment with IEA caused cytotoxicity and arrested cells in the sub-G0/G1 phase. Subsequent assays confirmed apoptosis-mediated cell death with mitochondrial membrane depolarization and alleviation of oxidative stress. IEA abrogates IL-6, VEGF, angiopoietin-2, and alters Th1/Th2 cytokines. IEA potentiated the effect of imatinib even at lower doses by affecting FAK/STAT/Akt/ERK pathways. IEA possesses antitumor potential against CML and increases the efficacy of imatinib when used in combination, suggesting utilization of IEA as an adjuvant therapy for better management of CML in the future.

The lncRNA ZNF667-AS1 Inhibits Propagation, Invasion, and Angiogenesis of Gastric Cancer by Silencing the Expression of N-Cadherin and VEGFA.

Purpose Gastric cancer is one of the most common malignancies with high mortality worldwide. It is known that long noncoding RNAs (lncRNAs) play important roles in the pathogenesis of gastric cancer. This study investigates the role of lncRNA ZNF667-AS1 in gastric cancer cells. Methods We have applied real-time quantitative PCR (qPCR) to study the levels of ZNF667-AS1 in gastric cancer biopsies and cell lines. The effects of ZNF667-AS1 on the propagation, clonogenicity, metastasis, and angiogenesis of gastric cancer cells were evaluated by calorimetry, colony formation, cell migration, and angiogenesis assays. Western blotting was used to identify the levels of proteins involved in cancer invasion and angiogenesis signaling pathways. Result It was found that lncRNA ZNF667-AS1 was downregulated in gastric cancer biopsies. Overexpression of ZNF667-AS1 reduced the propagation, migration, and angiogenesis of gastric cancer cells. Molecular mechanism studies displayed that the high level of lncRNA ZNF667-AS1 promoted the expression of E-cadherin and inhibited the expression of N-cadherin and VEGFA, leading to the inhibition of the proliferation, migration, and angiogenesis of gastric cancer cells. Conclusion As a tumor suppressor gene, lncRNA ZNF667-AS1 significantly hinders the propagation, metastasis, and angiogenesis of gastric cancer cells by promoting the expression of E-cadherin and inhibiting the expression of N-cadherin and VEGFA. Therefore, lncRNA ZNF667-AS1 could play a synergistic therapeutic role by targeting tumor cells and vascular endothelial cells, which represents a new therapeutic scheme for novel therapeutics of gastric cancer.

Available methods to enhance regenerative potential of plastic materials for bone defects replacement in orthopedics. Part 2. Use of autologous human platelet lysate

In the previous article, we talked about the use of platelet-rich plasma. One of the promising ways to stimulate the processes of repair and regeneration in the tissues of the damaged organ in different types of pathology is the use of platelet-rich plasma lysate. This part of the literature review covers the mechanism of action of platelet-rich plasma lysate, indications and contraindications for its use, describes the results of treatment when platelet-rich plasma lysate is used to stimulate osteogenesis. The preparation technology provides for the removal of all cellular components from the plasma, so it becomes possible to store the obtained graft for a long time. The procedure for the preparation of platelet lysate allows the simultaneous isolation of all growth factors from the cells, since the platelet lysis occurs. Lysate of platelet concentrates can be considered as a preparation that contains a complete set of stimulating growth factors. Under the influence of the lysate, the proliferation of latent osteoblasts is resumed, the signaling pathways of angiogenesis are activated, the secretion of the factors accelerating angiogenesis is stimulated, the differentiation of osteoblasts and the formation of bone tissue are triggered. The aim of this article is to summarize the results of treatment using autologous platelet lysate to improve bone regenerative potential in orthopaedics. In a final article, we shall look at the ways to use autologous red bone marrow.

Transcriptomic analysis of the chorioallantois of equine cloned pregnancies

Despite notable accomplishments over the last 25 years, somatic cell nuclear transfer (SCNT) still suffers from high pregnancy loss rate and a high incidence of abnormal offspring. The problems could be related to placental anomalies, which in turn may originate in defective epigenetic reprogramming. It is well known that a high percentage of SCNT pregnancies are associated with placental abnormalities including hydrallantois, reduced vascularization, and edematous placental membranes. To date, there is little known about the underlying mechanisms of the placental pathologies associated with cloned equine fetuses. Our study aimed to (1) characterize the transcriptome and biological pathways of placentas from cloned pregnancies at term, and (2) compare the transcriptome of placentas from cloned pregnancies that result in normal foals with placentas of cloned pregnancies resulting in non-viable foals. Chorioallantois (CA) was collected from the pregnant horn of placentas after normal parturition (mean gestational age of 339 days). A total of 19 samples were analyzed from 5 healthy, age-matched control pregnancies by artificial insemination (CON), 7 cloned pregnancies resulting in a healthy live foal (SCNT-L), and 7 cloned pregnancies resulting in a foal that died within the hour after birth (SCNT-D). Total RNA was extracted and sequenced, generating ∼33 million reads/sample. The raw reads were trimmed (Trimmgalore) and mapped (STAR) to the current equine reference genome. Mapped reads were quantified (FeatureCounts), and differentially expressed genes (DEGs) were evaluatedusing DESeq2, followed by gene ontology (GO) analysis. Macroscopically, 50% of the placentas from SCNT derived pregnancies showed edema. We identified 1332 DEGs (FDR <0.05, FoldChange>2) between the SCNT-L and CON groups and 1651 DEGs between the SCNT-D and CON groups. Pathway analysis of DEGs demonstrated that these genes are involved with extracellular matrix/ structure organization, response to growth factor, angiogenesis, and integrin signaling pathway independent of the outcome of the cloned pregnancies. When comparing the SCNT-L and SCNT-D groups, we identified 44 DEGs, mainly involved in inflammatory processes, even though histopathological examinations of the CA samples did not reveal any ongoing inflammatory process. Overall, our data highlights the underlying molecular pathways associated with abnormal placental development in cloned equine fetuses. Moreover, our data demonstrated the disruption of angiogenesis in the cloned placenta, manifested as excessive edema in the placenta. Similarly, in a previous study, we demonstrated the association between abnormal angiogenesis and placental edema and hydrallantois in equine pregnancy, highlighting the importance of adequate angiogenesis in placental function. This study was funded by the Center for Equine Health at UC Davis, FAPESP 2020/10260-3, the BOF Special Research Fund at Ghent University, a Fellowship of the Belgian American Educational Foundation, and the L'Oréal-UNESCO Fellowship.

Biological and Clinicopathological Characteristics of OPN in Cervical Cancers.

Background: Cervical cancer (CC) is the most common gynecological malignancy. Recently, an increasing number of studies have indicated that osteopontin (OPN) is a promising diagnostic and prognostic biomarker for CC. However, the biological role and detailed mechanism of OPN in CC remain unclear. Methods: The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets and a clinical sample microarray were used in our study. To identify the clinicopathological characteristics of OPN in CC, we compared the expression of OPN between normal and CC tissue samples and analyzed the correlations between OPN expression and multiple clinicopathological features. To identify biological processes involving OPN, OPN-associated genes were screened with Pearson correlation analysis and applied in hallmark gene set enrichment analysis (GSEA). Additionally, leukocyte infiltration was evaluated based on OPN expression. Finally, OPN-related signaling pathways were identified by GSEA. Results: OPN expression was higher in CC samples than in normal tissue samples and positively correlated with age, FIGO stage, tumor size, lymphovascular invasion and an unfavorable prognosis. OPN-associated genes were mainly enriched in the immune response, and increased OPN expression was accompanied by increased M2 macrophage infiltration. Additionally, OPN was correlated with hypoxia, high glycolytic metabolism, apoptosis, angiogenesis, epithelial-mesenchymal transition and multiple signaling pathways (the p53 pathway, the PI3K/Akt pathway, IL6/STAT3 signaling, mTORC1 signaling and KRAS signaling). Conclusion: Our study showed that OPN is involved in immunological activities and multiple tumor processes, identifying it as a potential therapeutic target and useful prognostic factor in CC patients.

Selected Articles from This Issue

Interleukin-15 (IL-15) regulates the generation, persistence, and differentiation of natural killer (NK) and CD8 T cells. To expand its role in cancer immunotherapy, Chen and colleagues designed a multitumor model treated with the combination of intratumoral anti-CD40, parenteral IL-15 and anti-PD-1. Anti-CD40 given intratumorally not only reduced dosing requirements and toxicity, but it also stimulated the antitumor activity of the distal tumors, suggestive of an abscopal effect. IL-15 with intratumoral anti-CD40 was additive/synergistic, and the inclusion of the anti-PD-1 was additive to the combination. Thus, this treatment shows promise for patients with solid tumors or resistance to anti-PD-1 therapy.Cancer-associated fibroblasts (CAF) are a heterogenous cell population with a complex immunoregulatory role, which has not been well defined in head and neck squamous cell carcinoma (HNSCC). Leveraging protein activity inference tools to analyze scRNA-sequencing of HNSCC patients, pre- and post-nivolumab, Obradovic and colleagues identified distinct subpopulations of head and neck CAF (HNCAF). Enrichment of a T-cell stimulating population (tsCAF) was found to associate with treatment response and ex vivo assays confirmed this CAF subset reduces CD8 T-cell exhaustion. Analysis of an independent HNSCC cohort treated with pembrolizumab validated this tsCAF subset as a potential biomarker of response.Approximately 30% of MSI-H/dMMR tumors do not respond to immune checkpoint inhibitors (ICI), highlighting the importance of identifying predictive biomarkers or molecular profiling correlated with resistance to these agents. Here, Chida and colleagues performed transcriptomic profiling of MSI-H/dMMR gastrointestinal tumors treated with PD-1 blockade. The study revealed that nonresponders had the enrichment of epithelial mesenchymal transition, angiogenesis, hypoxia, mTORC1, KRAS, Wnt/β-catenin, TGF-β, and various metabolism-related signaling pathways, which have been associated with an immunosuppressive tumor microenvironment. Meanwhile, the IFN-γ pathway was upregulated in responders. High expression of VEGF-A might predict a negative response to ICI in this population.Acral melanoma is a rare subtype of melanoma that arises on non-sun exposed skin of the hands, feet, and nailbeds. Patients with acral melanoma have lower response rates to immunotherapy than those with nonacral cutaneous melanoma, suggesting unique microenvironmental features. In their study, Li and colleagues performed comprehensive single cell analysis of a cohort of acral melanoma specimens and found them to harbor fewer effector CD8 T cells and NK cells and a complete absence of γδ T cells compared to non-acral cutaneous melanoma. VISTA, TIGIT, and adenosine signaling were identified as targetable immune checkpoints in acral melanoma.

Clinical, genomic, and transcriptomic correlates of response to immune checkpoint blockade-based therapy in a cohort of patients with angiosarcoma treated at a single center

BackgroundAngiosarcoma is a histologically and molecularly heterogeneous vascular neoplasm with aggressive clinical behavior. Emerging data suggests that immune checkpoint blockade (ICB) is efficacious against some angiosarcomas, particularly cutaneous angiosarcoma of...

Multigene Panel Sequencing Reveals Cancer-Specific and Common Somatic Mutations in Colorectal Cancer Patients: An Egyptian Experience.

This study aims at identifying common pathogenic somatic mutations at different stages of colorectal carcinogenesis in Egyptian patients. Our cohort included colonoscopic biopsies collected from 120 patients: 20 biopsies from patients with inflammatory bowel disease, 38 from colonic polyp patients, and 62 from patients with colorectal cancer. On top of this, the cohort included 20 biopsies from patients with non-specific mild to moderated colitis. Targeted DNA sequencing using a customized gene panel of 96 colorectal related genes running on the Ion Torrent NGS technology was used to process the samples. Our results revealed that 69% of all cases harbored at least one somatic mutation. Fifty-seven genes were found to carry 232 somatic non-synonymous variants. The most frequently pathogenic somatic mutations were localized in TP53, APC, KRAS, and PIK3CA. In total, 16 somatic mutations were detected in the CRC group and in either the IBD or CP group. In addition, our data showed that 51% of total somatic variants were CRC-specific variants. The average number of CRC-specific variants per sample is 2.4. The top genes carrying CRC-specific mutations are APC, TP53, PIK3CA, FBXW7, ATM, and SMAD4. It seems obvious that TP53 and APC genes were the most affected genes with somatic mutations in all groups. Of interest, 85% and 28% of the APC and TP53 deleterious somatic mutations were located in Exon 14 and Exon 3, respectively. Besides, 37% and 28% of the total somatic mutations identified in APC and TP53 were CRC-specific variants, respectively. Moreover, we identified that, in 29 somatic mutations in 21 genes, their association with CRC patients was unprecedented. Ten detected variants were likely to be novel: six in PIK3CA and four variants in FBXW7. The detected P53, Wnt/βcatenin, Angiogenesis, EGFR, TGF-β and Interleukin signaling pathways were the most altered pathways in 22%, 16%, 12%, 10%, 9% and 9% of the CRC patients, respectively. These results would contribute to a better understanding of the colorectal cancer and in introducing personalized therapies for Egyptian CRC patients.

Novel Angiogenic Regulators and Anti-Angiogenesis Drugs Targeting Angiogenesis Signaling Pathways: Perspectives for Targeting Angiogenesis in Lung Cancer.

Lung cancer growth is dependent on angiogenesis. In recent years, angiogenesis inhibitors have attracted more and more attention as potential lung cancer treatments. Current anti-angiogenic drugs targeting VEGF or receptor tyrosine kinases mainly inhibit tumor growth by reducing angiogenesis and blocking the energy supply of lung cancer cells. However, these drugs have limited efficiency, raising concerns about limited scope of action and mechanisms of patient resistance to existing drugs. Therefore, current basic research on angiogenic regulators has focused more on screening carcinogenic/anticancer genes, miRNAs, lncRNAs, proteins and other biomolecules capable of regulating the expression of specific targets in angiogenesis signaling pathways. In addition, new uses for existing drugs and new drug delivery systems have received increasing attention. In our article, we analyze the application status and research hotspots of angiogenesis inhibitors in lung cancer treatment as a reference for subsequent mechanistic research and drug development.

Transcriptomic Profiling of MSI-H/dMMR Gastrointestinal Tumors to Identify Determinants of Responsiveness to Anti-PD-1 Therapy.

Transcriptomic profiling was performed for microsatellite instability-high (MSI-H)/mismatch repair-deficient (dMMR) gastrointestinal tumors to determine the predictors of response to PD-1 blockade. Thirty-six patients with MSI-H/dMMR gastrointestinal tumors, including gastric cancer, colorectal cancer, cholangiocarcinoma, small intestine cancer, and pancreatic cancer, being treated with PD-1 blockade were analyzed. We conducted the transcriptomic analysis of gastrointestinal tumors using RNA sequencing data, including the consensus molecular subtypes (CMS) of colorectal cancer. Gene set enrichment analysis (GSEA) demonstrated that non-responders had upregulations of epithelial-mesenchymal transition, angiogenesis, hypoxia, mTORC1, TNF-α, KRAS, Wnt/β-catenin, TGF-β, and various metabolism-related signaling pathways. Meanwhile, the IFNγ pathway was enriched in responders. On the basis of the leading-edge analysis of GSEA, VEGF-A was significantly correlated with enriched pathways in non-responders. Patients with high VEGF-A expression, compared with those with low expression, had significantly shorter progression-free survival [PFS; median 4.8 months vs. not reached (NR), P = 0.032] and overall survival (median 11.1 months vs. NR, P = 0.045). Among 13 patients with colorectal cancer evaluable for CMS classification, the objective response rate was 100%, 0%, 0%, and 16.7% in CMS1, CMS2, CMS3, and CMS4, respectively. Patients with CMS1 had significantly longer PFS (NR vs. 4.8 months, P = 0.017) than those with CMS2, CMS3, or CMS4. Several transcriptomic features, including CMS classification and related genes, were associated with response to PD-1 blockade in MSI-H/dMMR gastrointestinal tumors. These findings can help develop predictive biomarkers or combination immunotherapies.

Investigation of the Effect of High-Intensity Interval Training (HIIT) on the Expression of the Genes miR-126, miR-296, HGS and VEGF-A Protein Levels in Tumor Tissue in Female Mice

Background &amp; Objective: The studies of the last two decades have shown that regular training is associated with the reduction of mortality among patients with breast cancer, and has an important role in inhibiting breast cancer progression as well as treatment of the disease. However, the micro-molecular mechanisms involved are not fully understood yet. However, the aim of the present study was to investigate the effect of a high-intensity interval training (HIIT) on the expression of miR-126, miR-296, HGS gene, and VEGF-A protein level in tumor tissue of the mice with breast cancer. &#x0D; Materials &amp; Methods: For this purpose, 12 BALB/c mice (6-8 weeks, weight 19 ± 1.05 g), after induction of cancer (MC4-L2 subcutaneous injection into the right side of the mice), were randomly divided into two groups with 6 in each group: HIIT and control. Each session of HIIT group program includes six intervals of three minutes and 20 seconds with an intensity of 85 to 90 percent of VO2max and 1-minute recovery with 30 to 35 percent VO2max intensity between each interval; and the animals were sacrificed 24 hours after the last training session and the expression of miR-126, miR-296, HBS gene and VEGF protein levels in tumor tissues were examined.&#x0D; Results: The statistical results showed that the implementation of HIIT significantly reduced the expression of miR-296 and subsequently increased the HGS gene expression and led to increased expression levels of miR-126 and decreased levels of VEGF-A protein in the training group compared to the control group (p&lt;0.05).&#x0D; Conclusion: It seems that HIIT, by suppressing tumor angiogenesis signaling pathways, could be an effective intervention to inhibit the growth of breast tumors.