Peritumoral Expression Research Articles

Serum amyloid A promotes glycolysis of neutrophils during PD-1 blockade resistance in hepatocellular carcinoma

The response to programmed death-1 (PD-1) blockade varies in hepatocellular carcinoma (HCC). We utilize a panel of 16 serum factors to show that a circulating level of serum amyloid A (SAA) > 20.0 mg/L has the highest accuracy in predicting anti-PD-1 resistance in HCC. Further experiments show a correlation between peritumoral SAA expression and circulating SAA levels in patients with progressive disease after PD-1 inhibition. In vitro experiments demonstrate that SAA induces neutrophils to express PD-L1 through glycolytic activation via an LDHA/STAT3 pathway and to release oncostatin M, thereby attenuating cytotoxic T cell function. In vivo, genetic or pharmacological inhibition of STAT3 or SAA eliminates neutrophil-mediated immunosuppression and enhances antitumor efficacy of anti-PD-1 treatment. This study indicates that SAA may be a critical inflammatory cytokine implicated in anti-PD-1 resistance in HCC. Targeting SAA-induced PD-L1+ neutrophils through STAT3 or SAA inhibition may present a potential approach for overcoming anti-PD1 resistance.

Proportion and distribution of PD-L1 expression and associated clinicopathologic features in patients with microsatellite instability-high gastric cancer.

387 Background: The frequency of microsatellite instability-high (MSI-H) phenotype in sporadic gastric cancers (GCs) widely varies from 8% to 20%. From prior data, MSI-H phenotypes are significantly associated with female sex, older age, antral location of the tumor, well to moderate differentiation, intestinal type, non-signet ring cell component, mucinous histologic type, a moderate to severe lymphoid stromal reaction, and a lower TNM stage. This study investigated the proportion and distribution of PD-L1 expression of MSI-H GCs compared to microsatellite stable (MSS) GCs. Methods:The study consists of 847 cases of unselected GC. MSI analysis was performed on all GC cases by immunohistochemistry for mismatch repair proteins followed by multiplex polymerase chain reaction. Among them, 72 GCs were found to be an MSI-H phenotype. After matching the T category of GCs, 200 cases of GC with MSS phenotype were selected for a control group. Immunohistochemistry for PD-L1 (SP263) was performed on 72 MSI-H GCs and 200 MSS GCs. Results: Patients with MSI-GC were significantly associated with advanced age (p <0.001), advanced tumor (p=0.0173), antral location of the tumor(p=0.0018), and T2 and T3 category (p=0.0031) but not T1 or T4 category. Histologically, MSI-H GCs occurred frequently in ulcerofungating mass (45.8%, p < 0.0001)), intestinal type (73.6%, p=0.0102), well to moderately differentiated tumor (56.9%, p=0.0102), tubular histologic type (66.7%, p=0.0013), tumor necrosis (18.1%, p <0.0001), and peritumoral neutrophilic infiltrate (40.3%, p <0.001) compared to those of 200 MSS GCs. In the evaluation of PD-L1, 55 (76.4%) MSI-H tumors revealed PD-L1 expression with ≥ 1 CPS. The predominant PD-L1 staining was found in intratumoral and peritumoral immune cells (66.7%) than within tumor cells (9.7%). The distribution of positive immune cells was predominantly located in the periphery of the tumor (37.5%) than within the tumor (9.7%). According to the criteria for PD-L1 positivity as at least 10 CPS, approximately 50% of MSI-H GCs were considered PD-L1 positive tumors with at least 10 CPS. Conclusions: In this study, the prevalence of MSI-H phenotype is approximately 8.5% of unselected GCs. Approximately 76% of patients with MSI-H tumors showed PD-L1 expression with at least 1 CPS, and 50% revealed PD-L1 expression with at least 10 CPS, who can respond better to immune checkpoint inhibitors with promising therapeutic benefits. Due to the peritumoral expression of immune cells in PD-L1 positive GCs with MSI-H phenotype, different endoscopic approaches to obtain tumor tissues for PD-L1 evaluation would be considered, such as sampling tissues at the tumor periphery.

LncRNA XIST Interacts with Regulatory T Cells within the Tumor Microenvironment in Chronic Hepatitis B-Associated Hepatocellular Carcinoma.

Alterations in the expression of several long non-coding RNAs (lncRNAs) have been shown in chronic hepatitis B-associated hepatocellular carcinoma (CHB-HCC). Here, we aimed to investigate the association between the expression of inflammation-associated lncRNA X-inactive specific transcript (XIST) and the type of inflammatory cells within the tumor microenvironment. Twenty-one consecutive cirrhotic patients with CHB-HCC were included. XIST expression levels were investigated on formalin-fixed paraffin-embedded (FFPE) tumoral and peritumoral tissue samples by real-time polymerase chain reaction (RT-PCR). Immunohistochemical staining for CD3, CD4, CD8, CD25, CD163, CTLA4, and PD-1 were performed. The findings were statistically analyzed. Of the 21 cases, 11 (52.4%) had tumoral and 10 (47.6%) had peritumoral XIST expression. No significant association was found between the degree of inflammation and XIST expression. The number of intratumoral CD3, CD4, CD8 and CD20 positive cells was higher in XIST-expressing tumors, albeit without statistical significance. Tumoral and peritumoral XIST expression tended to be more common in patients with tumoral and peritumoral CD4high inflammation. The number of intratumoral CD25 positive cells was significantly higher in XIST-expressing tumors (p=0.01). Tumoral XIST expression was significantly more common in intratumoral CD25high cases (p=0.04). Peritumoral XIST expression was also more common among patients with CD25high peritumoral inflammation, albeit without statistical significance (p=0.19). lncRNA XIST is expressed in CHB-HCC and its expression is significantly associated with the inflammatory tumor microenvironment, particularly with the presence and number of CD25 (+) regulatory T cells. In vitro studies are needed to explore the detailed mechanism.

Peritumoral SPARC expression induced by exosomes from nasopharyngeal carcinoma infected Epstein-Barr virus: A poor prognostic marker.

Epstein-Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC) cells have high metastatic potential. Recent research has revealed that the interaction of between tumor cells and the surrounding stroma plays an important role in tumor invasion and metastasis. In this study, we showed the prognostic value of expression of SPARC, an extracellular matrix protein with multiple cellular functions, in normal adjacent tissues (NAT) surrounding NPC. In the immunohistochemical analysis of 51 NPC biopsy specimens, SPARC expression levels were significantly elevated in the NAT of EBER (EBV-encoded small RNA)-positive NPC compared to that in the NAT of EBER-negative NPC. Moreover, increased SPARC expression in NAT was associated with a worsening of overall survival. The enrichment analysis of RNA-seq of publicly available NPC and NAT surrounding NPC data showed that high SPARC expression in NPC was associated with epithelial mesenchymal transition promotion, and there was a dynamic change in the gene expression profile associated with interference of cellular proliferation in NAT, including SPARC expression. Furthermore, EBV-positive NPC cells induce SPARC expression in normal nasopharyngeal cells via exosomes. Induction of SPARC in cancer-surrounding NAT cells reduced intercellular adhesion in normal nasopharyngeal structures and promoted cell competition between cancer cells and normal epithelial cells. These results suggest that epithelial cells loosen their own binding with the extracellular matrix as well as stromal cells, facilitating the invasion of tumor cells into the adjacent stroma by activating cell competition. Our findings reveal a new mechanism by which EBV creates a pro-metastatic microenvironment by upregulating SPARC expression in NPC.

Hepatocellular carcinoma: tumor heterogeneity and recurrence after preoperative locoregional therapy.

How locoregional therapy (LRT) may change tumor compositional and functional heterogeneity, consequently contributing to treatment resistance and tumor recurrence remains largely unknown. A series of hepatocellular carcinomas (HCCs) treated with preoperative locoregional therapy (LRT) that relapsed after surgery was studied. Thirty HCCs comprising 15 patients treated with LRT prior to liver transplantation (n = 14)/surgical resection (n = 1) were studied. Five patients undergoing pre-transplant LRT, comprising 11 HCCs, presented tumor recurrence (median recurrence time = 10months). Clinicopathological data and immunoexpression of proliferation markers (Ki67, p53), cholangiocytic/hepatic progenitor cell (HPC) markers (EpCAM)/BerEp4, CK19, CK7) and mesenchymal markers (ASMA, vimentin) were evaluated in tumoral epithelial/stromal cells and in peritumoral parenchyma. Higher grading of tumor differentiation, microvascular invasion and tumoral cell p53 expression significantly associated with recurrence (p < 0.05). Piecemeal necrosis features were more frequent in tumors that recurred (p < 0,05). Tumoral and peritumoral Ki67 expression and EpCAM, CK19 and CK7 expression in tumoral cells tended to be higher in treated tumors that recurred. Peritumoral expression of cholangiocytic/HPC markers and tumoral epithelial and stromal cellular expression of mesenchymal markers tended to be higher for tumors without recurrence. Recurrence after transplant with preoperative LRT might be associated with poor HCC differentiation, higher cellular proliferation rate, peritumoral piecemeal necrosis features and cholangiocytic/HPC phenotypes. Understanding HCC progression factors after LRT might be important for optimizing patient selection for treatment, improving surveillance after LRT and to explore synergies between LRT and systemic targeted therapies to prevent recurrence.

Clinical/prognostic significance of Syndecan-1 expression in invasive breast carcinoma with distant metastasis and its correlation with tumor immunity

ObjectiveBreast Cancer (BC) is the most common malignant tumor for women in the world. 90% of BC-associated deaths are attributed to distant metastasis (DM). Therefore, there is an urgent need for a novel molecular target for the treatment of distant metastatic breast cancer (DMBC). Syndecan-1 (SDC-1) is a cell surface heparan sulfate proteoglycan (HSPG). This study aims to study the expression patterns of SDC-1 in invasive breast carcinoma (IBC) with DM and to analyze its relationship with different clinicopathologic features, stromal tumor infiltrating lymphocytes (sTILs) status and the clinical outcomes. MethodsA total of 50 DM breast cancer and 100 non-distant metastasis (non-DM) breast cancer patients in West China Hospital, Sichuan University from January 1, 2011 to December 31, 2011 were collected. Immunohistochemical (IHC) method was used to detect the expression of SDC-1, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor-2 (HER2), and Ki-67 in 150 specimens of patients with IBC. STILs were used to evaluate immune cells in the stromal tissue within the tumor. Various clinicopathologic characteristics were retrospectively analyzed, and follow-up information were collected for prognosis analyses. The expression pattern difference of SDC-1 in the DM group and the non-DM group and its correlation with clinicopathologic characteristics of IBC were analyzed. ResultsCompared with the non-DM group, SDC-1 had higher cytoplasmic (90.0%) and stromal diffuse (70.0%) expressions and lower stromal peritumoral (18.0%) expression in the DM group. SDC-1 cytoplasmic expression was significantly associated with HER2-positive and high Ki-67 index in DM group, and with high histological grade and lymph node (LN) metastasis in non-DM group (P < 0.05). Compared with the non-DM group, the membranous expression of SDC-1 in the DM group was related to higher histological grade and T stage, higher frequency of LN involvement. Meanwhile, the expression pattern of SDC-1 in tumor stroma was associated with sTILs status (P < 0.05). The different combinations of SDC-1 staining patterns were correlated with clinicopathological features, biomarkers and sTILs status between DM group and non-DM group.There was no significant difference in overall survival between DMBC with different expression patterns of SDC-1. ConclusionThe cytoplasmic and stromal expressions of SDC-1 in the primary lesion of IBC are closely associated with DM, and the stromal expression of SDC-1 is correlated with tumor immune microenvironment. SDC-1 is expected to be a potential new marker for predicting the risk of DM in IBC.

Abstract 4588: Vascular regulation of T-cell exclusion in melanoma

Abstract T-cell position in the tumor microenvironment determines the probability of target encounter and tumor killing. CD8+ T-cell exclusion from the tumor parenchyma is associated with poor response to immunotherapy and yet the biology that underpins this distinct pattern remains unclear. Here we show that the vascular destabilizing factor angiopoietin-2 (ANGPT2) causes compromised vascular integrity in the tumor periphery, leading to impaired T-cell infiltration within tumor. We analyzed the spatial regulation of ANGPT2 in whole tumor cross-sections and its impact on CD8+ T-cell position, vascular integrity, and response to immunotherapy using syngeneic murine melanomas. We found that T-cell exclusion was associated with peritumoral expression of ANGPT2 and elevated vascular leakage in human and murine melanomas. Both pharmacological and genetic blockade of ANGPT2 promoted CD8+ T-cell infiltration to the tumor core, exerting antitumor effects. Importantly, the reversal of T-cell exclusion following ANGPT2 blockade not only enhanced response to anti-PD-1 therapy in immunogenic, therapy responsive mouse melanomas, but also rendered non-responsive tumors susceptible. Therapeutic response after ANGPT2 blockade, driven by improved CD8+ T-cell infiltration, coincided with TIE2 signaling activation and increased vascular integrity in the tumor periphery, indicating that maintenance of vascular integrity within tumors is necessary for robust response to immunotherapy. These data highlight ANGPT2/TIE2 signaling as a key mediator of T-cell exclusion and a promising target to potentiate checkpoint inhibitor efficacy in melanoma. Citation Format: Ha-Ram Park, Anahita Shiva, Portia Cummings, Richard Carvajal, Gavin Thurston, Joon-Yong An, Amanda W. Lund, Hee Won Yang, Minah Kim. Vascular regulation of T-cell exclusion in melanoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4588.

Immune Microenvironment of Muscular-Invasive Urothelial Carcinoma: The Link to Tumor Immune Cycle and Prognosis.

In this study, we investigated the relationship between the tumor immune microenvironment (TIME), histological differentiation and hypoxia in patients with muscular-invasive urothelial carcinomas (MIUC) after radical cystectomy. Forty-two cases of pT2-3N0M0 MIUCs underwent clinical, histological and immunohistochemical evaluation by counting CD8+, FOXP3+, CD68+, CD163+ cells and polymorphonuclear leukocytes (PMN) in intra-tumoral and peritumoral areas, assessing PD-L1 and GLUT1 expression for defining the impact of tumor immune contexture on patients’ outcomes. Five-year survival rates and overall survival were calculated. Most of the MIUCs demonstrated the immune-desert or immune-excluded TIME, reflecting altered mechanisms of T-cells’ activation or traffic into tumors. Tumor immune contexture was closely related to histological differentiation. CD8+ cells were scant in MIUCs with papillary and squamous differentiation, while basal-like or mesenchymal-like histological differentiation was associated with increased density of CD8+ cells. A high rate of PD-L1 expression (47.6%) was not related to immune cell infiltration. M2-macrophages predominated under CD8+ lymphocytes. The abundance of PMN and CD163+ macrophages in MIUCs was associated with high GLUT1 expression. CD8+, CD68+, FOXP3+ cells and PD-L1 status did not affect patients’ outcomes, while high CD163+ density and PMN infiltration were associated with the unfavorable outcome of patients with MIUC. These data drive the hypothesis that in MIUC, immune escape mechanisms are shifted towards the role of the innate immunity cells rather than CD8+ lymphocytes’ functioning.

Association between Foxp3 Tumor Infiltrating Lymphocyte Expression and Response After Chemoradiation in Nasopharyngeal Carcinoma

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a carcinoma originating from the surface epithelium of the nasopharynx with the highest incidence in China and South East Asia. Currently, many researchers are developing tumor microenvironment which can be assessed by tumor-infiltrating lymphochyte, and its association with treatment response in several tumors, including NPC. Foxp3, known as a regulatory T cell (Treg) marker, plays a role in the immunoregulatory environment of tumor cells and can be used as a prognostic factor. The relationship between Foxp3 expression and treatment response is considered as one of the factors affecting the prognosis of NPC. AIM: This study aims to determine the relationship between Foxp3 expression and treatment response in NPC. MATERIALS AND METHODS: A cross-sectional study was done to analyze the association between Foxp3 and treatment response in NPC. This study included 60 samples who were diagnosed with non-keratinizing NPC at the Department of Anatomical Pathology, Faculty of Medicine Universitas Indonesia/Cipto Mangunkusumo Hospital from January 2018 until December 2020. Immunohistochemistry was done to evaluate the expression of Foxp3. Foxp3 expression was evaluated in the intratumoral and peritumoral areas. RESULTS: Among 60 patients, the number of males were more than females (66.7%, 33.3%, respectively) with a ratio of 2:1. There was statistically significant difference between intratumoral and total Foxp3 expression and treatment response (p &lt; 0.05, p = 0.001, respectively); however, no significant differences found between peritumoral Foxp3 expression and treatment response (p = 0.114). CONCLUSION: Foxp3 expression had a statistically significant relationship with response therapy after chemoradiation.

PD-L1 Expression in Different Segments and Histological Types of Ovarian Cancer According to Lymphocytic Infiltrate.

Background and Objectives: Ovarian cancer is the leading cause of death among gynecological tumors. PD-1/PD-L1 immunoregulatory mechanism is activated in ovarian cancers. Lymphocyte infiltration is a significant factor that affects its expression. We analyzed the correlation between localization of lymphocytic infiltrate and PD-L1 expression in epithelial ovarian tumors. Materials and Methods: PD-L1 expression was analyzed in 328 subjects, 122 with epithelial ovarian carcinoma, 42 with atypical proliferative tumor, and 164 with benign epithelial ovarian tumor. Expression in central and invasive tumor parts in epithelial ovarian carcinoma was combined with the most pronounced lymphocyte reaction. Immunohistochemical analysis was performed using the tissue microarray and correlated with a set of histopathology parameters. Results: PD-L1 expression was most prominent in epithelial ovarian carcinoma with different levels of expression observed between invasive and central tumor segments. A high level of PD-L1 expression on tumor cells was more frequently present in the invasive than in the central tumor parts (p < 0.001) only in high-grade serous ovarian carcinoma (HGSC). There was no significant correlation between peritumoral lymphocytic infiltrate and PD-L1 expression regardless of tumor segment. In the central tumor parts of HGSC, there was a correlation of intratumoral lymphocytic infiltrate with a higher level of PD-L1 expression (p = 0.003). Conclusions: The most prominent PD-L1 expression was observed in the invasive tumor parts of HGSC. Only the central parts of the HGSC exhibited significant PD-L1 expression in association with considerable intratumoral lymphocytic infiltrate.

Correlation of peritumoral circWDR25 expression with the prognosis of patients with hepatocellular carcinoma after curative resection

To study the association between the expression of peritumoral circWDR25 (hsa-circRNA-0004310) secreted by hepatic stellate cells (HSCs) and the prognosis of the patients with hepatocellular carcinoma (HCC) after curative resection. HSCs cell line LX-2 was co-cultured separately with 3 liver cancer cell lines (Hep3B, SMMC-7721, and HCCLM3) in Transwell chambers to obtain tumor cell-activated HSCs. The supernatants of HSC cultures were collected to isolate the exosomes, from which total RNA was extracted to detect circRNA expression profile. We also collected specimens of paracancerous liver tissues from 288 HCC patients undergoing radical resection in our department from January, 2014 to October, 2015, and the expression levels of circWDR25 and α-SMA were detected with in situ hybridization. Log-rank test and Cox regression analysis were used for univariate and multivariate analysis of the factors affecting the patients' prognosis, respectively. Gene expression profiling revealed that the expression of circWDR25 was the most obviously up-regulated in the exosomes isolated from tumor-activated LX-2 cells. The expression of peritumoral circWDR25 was positively correlated with HSCs adjacent to the cancer loci (r=0.156, P=0.008). Multivariate analysis showed that a preoperative AST level >36 g/L, multiple tumors, a tumor diameter >5 cm, HSC>70, and circWDR25>190 were independent risk factors affecting the overall survival of HCC patients after radical resection; a preoperative AST level >36 g/L, multiple tumors, a tumor diameter >5 cm, presence of tumor thrombus, HSC>70, and circWDR25>190 were all independent risk factors for tumor-free survival in patients with liver cancer. Peritumoral circWDR25 and HSCs are factors affecting the prognosis of HCC patients after radical hepatectomy, and their high expression in the adjacent tissues is closely related to a poor prognosis of the patients.

HHLA2 Expression is Associated with Poor Survival in Patients with Hepatocellular Carcinoma.

BackgroundHuman endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) is a member of the B7 family; however, little is known regarding its expression and clinical relevance in hepatocellular carcinoma (HCC).MethodsTo better characterize HHLA2 expression in HCC, we analyzed its expression by in situ staining and further investigated its correlation with immune infiltration and patient prognosis.ResultsHHLA2 was primarily expressed in the peri-tumor region of HCC tissues and co-localized with CD68+ monocytes/macrophages. In vitro analysis and multi-immunofluorescence staining showed up-regulated HHLA2 expression in tumor-activated monocytes/macrophages, and HHLA2+ monocytes/macrophages expressed high levels of HLA-DR in HCC tissue. A correlation analysis showed that samples displaying high HHLA2 expression in the peri-tumor region had significant tumor infiltration of CD204+ and CD11b+ cells, and low expression of genes associated with an anti-tumor immune response. The high level of peri-tumoral HHLA2 expression was associated with a poor patient overall survival (OS; P = 0.008). A multivariate analysis revealed that HHLA2 expression in the peri-tumor region was an independent prognostic factor for OS (hazard ratio = 1.872, p = 0.003). Moreover, the expression of HHLA2 was negatively correlated with PD-L1, and patients exhibiting HHLA2 and programmed cell death-ligand 1(PD-L1) co-expression had the shortest survival time.ConclusionHHLA2 expression represented an immunosuppressive microenvironment in HCC, and may serve as a potential target for immunotherapy.

Inflammatory Biomarkers as Promising Predictors of Prognosis in Cervical Cancer Patients

Introduction: Increasing evidence demonstrates a crucial role of inflammation in inducing and promoting several cancers. Pro-inflammatory upregulation of cytokines such as IL-6 has been implicated in cervical cancer development and progression through several mechanisms, for example, by inducing platelet production, activation, and aggregation. The aim of the study was to evaluate the effective prognostic impact of inflammatory biomarkers such as platelet count, platelet to lymphocyte ratio (PLR), and IL-6 in cervical cancer patients. Materials and Methods: Between 2016 and 2019, 108 out of 159 patients with cervical cancer have been enrolled. Cutoff level of pretreatment platelet count and PLR was identified by using the ROC curve. IL-6 tumoral and peritumoral expression was analyzed and stratified as low and high (low expression: 0 and +1; marked expression: +2 and +3). Results: Median follow-up duration was 30 months (range 16–44). Patients with higher platelet counts showed worse DFS and OS (DFS p < 0.001; OS p < 0.001). Cumulative rates of DFS and OS in patients with lower PLR were higher than in patients with higher values of PLR (DFS p = 0.032; OS p < 0.001). Survival analysis showed a better prognosis in patients with lower IL-6 expression (DFS p < 0.001; OS p < 0.001). Conclusion: Nowadays, causal relationship between inflammation, innate immunity, and cancer is more widely accepted. However, many of the molecular and cellular mechanisms mediating this relationship remain unresolved. Ongoing inflammatory response was associated with poor outcomes in cervical cancer patients. A higher pretreatment platelet count and PLR value associated with higher IL-6 tumoral expression could be used to predict poor prognosis in cervical cancer patients.

Macrophages expressing TREM-1 are involved in the progression of HPV16-related oropharyngeal squamous cell carcinoma

Introduction Many types of research have been performed to improve the diagnosis, therapy, and prognosis of oropharyngeal carcinomas (OP-SCCs). Since they arise in lymphoid-rich areas and intense lymphocytic infiltration has been related to a better prognosis, a TREM-1 putative function in tumour progression and survival has been hypothesized. Materials and methods Twenty-seven human papillomavirus (HPV) 16+ OP-SCC specimens have been analyzed to relate TREM-1 expression with histiocytic and lymphocytic markers, HPV presence and patients’ outcome. Results No differences have been shown between intratumoral and stromal CD4+ cells, while intratumoral CD8+ lymphocytes are higher with respect to the tumour stroma (p = .0005). CD68+ cells are more than CD35+ and TREM-1+; their presence is related to CD35± and TREM-1± histiocytes (p = .005 and .026, respectively). Intratumoral CD4+ lymphocytes are higher in p16+ cases (11/27) than in p16− (p = .042); moreover, p16 positivity correlates to a better survival (p = .034). CD4+, CD8+ and CD35+ cells have no impact on survival, while CD68 expression heavily influences progression and bad outcome (p = .037). TREM-1 positivity also leads to worst overall survival (p = .001): peritumoral expression and death-cause relationship are always significant, particularly when the cause is OP-SCC (p = .000). Conclusion While p16 shows to better stratify HPV16+ patients’ outcome, TREM-1+ macrophages suggest their key importance in HPV-related OP-SCCs progression. KEY MESSAGES TREM-1 positivity correlates to the worst overall survival of HPV16-positive OPSCCs-affected patients. p16-positive HPV16 related OPSCCs patients have a better prognosis with respect to p16-negative ones.

Galectins-1, -3, -7, -8 and -9 as prognostic markers for survival in epithelial ovarian cancer: A systematic review and meta-analysis.

Galectins are a family of proteins that have recently emerged as regulators of cancer biology. To investigate the impact of peritumoral and tumoral galectin expression on ovarian cancer prognosis. We searched Medline, Cochrane, and EMBASE databases from inception until March 22, 2020. All studies correlating galectins and ovarian cancer prognosis were selected. The literature search presented 11 studies, which contained 1034 patients. Meta-analysis was performed with RevMan 5.3 software. Studies were stratified into two groups depending on the location of galectin expression (peritumoral stroma or nucleus/cytoplasm of tumor cells). Tumoral galectin-7 and galectin-9 expression was significantly associated with poor overall survival (odds ratio [OR] 2.06, 95% confidence interval [CI] 1.32-3.21, P=0.001; OR 1.71, 95% CI 1.27-2.30, P<0.001, respectively). The total effect of high tumoral expression of galectins in overall survival and progression-free survival was significant (OR 1.51, 95% CI 1.02-2.23, P=0.04; OR 2.76, 95% CI 1.73-4.40, P<0.001, respectively). Our results suggest that galectins are implicated in ovarian cancer prognosis; however, further research is needed to ascertain their actual importance as well as their diagnostic accuracy.

Canine osteosarcoma checkpoint expression correlates with metastasis and T-cell infiltrate

BackgroundImmune-targeted therapies are being successfully implemented into cancer clinical practice. In particular checkpoint inhibitors are employed to modulate the immune microenvironment of solid tumors. We sought to determine the expression of PD-L1, HVEM, and B7H3 in human and canine osteosarcoma, and correlate expression with clinical features and tumor infiltrating lymphocytes in naturally-occurring canine osteosarcoma. MethodsFlow cytometry was used to measure ligand surface expression of five human and three canine cell lines. Immunohistochemistry was utilized for expression of ligands and lymphocyte markers in thirty-seven treatment-naïve canine osteosarcoma patients. ResultsAll cell lines expressed all three ligands at variable levels in both species. Metastatic lesions were associated with higher expression of all three ligands in patient tumor samples. PD-L1 expression strongly correlated with B7H3 and HVEM expression, while HVEM and B7H3 were weakly correlated. Whereas peritumoral T-cell expression positively correlated with PD-L1 and HVEM tumor expression, the presence of T-cells intratumorally were rare. Furthermore, intratumor penetration by T-cells was greatest in metastatic lesions, despite log-fold increases in peritumoral T-cells.In summary, PD-L1, HVEM, and B7H3 are expressed in osteosarcoma, with metastatic disease lesions expressing higher levels. We show for the first time that these ligands expressed on osteosarcoma cells positively correlate with each other and the presence of peritumoral T cell infiltration. Furthermore, osteosarcoma appears to be an intratumoral immune desert with significant resistance to effector T cells. Multiple agents targeting checkpoints are in clinical practice, and may have immune modulating benefit in osteosarcoma.

Hepatic Artery Embolization Enhances Expression of Programmed Cell Death 1 Ligand 1 in an Orthotopic Rat Hepatocellular Carcinoma Model: In Vivo and in Vitro Experimentation

PurposeTo evaluate the effects of hepatic artery embolization (HAE) on the expression of programmed cell death 1 ligand 1 (PD-L1) in an orthotopic rat hepatocellular carcinoma (HCC) model. Materials and MethodsA rat HCC model was established in Sprague–Dawley rats with the RH7777 cell line. Six animals each were assigned to receive HAE or sham treatment. Liver tissues were harvested 24 h after the procedure. Immunohistochemistry (IHC) was used to compare expression of PD-L1 and hypoxia-inducible factor (HIF)–1α in the intratumoral and peritumoral regions and normal liver tissue. In vitro cell culture study was performed for 24 h under normoxic and hypoxic conditions, and protein expression of PD-L1 and HIF-1α and the effects of HIF-1α inhibitors were assessed. ResultsIHC showed that PD-L1– and HIF-1α–positive areas were significantly larger in the HAE group vs the sham group in intratumoral (P = .006 and P < .001, respectively) and peritumoral regions (both P < .001). The expression of PD-L1 positively correlated with HIF-1α expression in the intratumoral region (r2 = 0.551; P < .001). In vitro cell culture study revealed that protein expression of PD-L1 and HIF-1α were significantly higher when cells were incubated under hypoxic vs normoxic conditions (P = .028 and P = .010, respectively). PD-L1 expression was suppressed significantly when the HIF-1α inhibitor rapamycin was added to the culture medium (P = .024). ConclusionsHAE enhances intratumoral and peritumoral PD-L1 expression in a rat HCC model. The HIF-1α pathway is a possible mechanism underlying increased intratumoral PD-L1 expression after HAE.

Brain metastasis PD-L1 and CD8 expression is dependent on primary tumor type and its PD-L1 and CD8 status.

BackgroundBrain metastases (Bmets) are frequent; however, limited data exist on the efficacy of immunotherapy in these lesions. The aims of the study were to analyze the immunohistochemical expressions of programmed death ligand 1 (PD-L1) and CD8 in Bmets and to compare them with their expressions in paired primary tumors, as well as correlate the results with clinicopathological features.MethodsThis is a retrospective study of 233 patients with Bmets and 111 paired primaries. Clinical, histological, and molecular data were recorded and compared with the immunohistochemical results of PD-L1 and CD8 expressions. The statistical analysis included χ2 test, Cramer’s V test, factorial analyses of variance, simple regression analysis, and Kaplan-Meier analysis with log-rank product limit estimation.ResultsPD-L1 expression was found in 23.6% of Bmets and in 29.0% of primary tumors with concordant expression between them in 75.5% of cases. Bmets PD-L1 expression was associated with primary tumor PD-L1 expression and the primary tumor type. Significant CD8 peritumoral expression was found in 68.6% of Bmets and in 87.7% of primary tumors. CD8 expression was concordant between primary and metastatic tumors in 73.3% of cases. Bmets CD8 expression was associated with primary tumor CD8 expression and primary tumor type. PD-L1 expression was associated with CD8 expression in both primary and metastatic tumors. The concordance between primary and metastatic tumor PD-L1 expression was independent of all factors studied. The concordance between primary and metastatic CD8 expressions was marginally associated to the time of Bmets development. No prognostic role for PD-L1 and CD8 expression in Bmets was found.ConclusionPD-L1 and CD8 Bmets expressions are associated with the primary tumor type and its PD-L1 and CD8 expressions. No factor predicts the discordance for PD-L1 expression, while time to Bmets development is associated with CD8 expression discordance.

Peritumoral Lymphangiogenesis in Breast Carcinoma

Objective: to assess the correlation between microscopically discernible peritumoral lymphangiogenesis and breast carcinoma survival rates; to reveal the regularities of macroscopic changes in the structure of axillary lymphatic system in breast cancer. Materials and Methods: We carried out morphometric examination of axillary tissue complexes that had been taken from 100 women during surgery for breast carcinoma, stage T1-3N1-2M0 and had been treated with sonolipodistruction. The density of realized lymphatic vessels was determined by counting the number of lymphangiogenesis "points". An anastomosis between individual lymphatic vessels was taken as a "point". Correlation of macroscopically distinguishable peritumoral lymphangiogenesis expression and survival rate was determined using Pearson's parametric analysis. Results: The analysis of lymphatic maps allowed us to reveal some patterns of axillary lymphatic collector reorganization in breast carcinoma and to distinguish the following types: "Sequential", "Magistral", "Uniform", "Radial" and "Reticulate". Based on the results of prospective observation, 16 patients from the observation group died within the period of 5 years. According to the Pearson correlation analysis, the correlation coefficient was 0,87, the determination coefficient was 0,7488, the value of p = 0,000015, which indicates that in the breast carcinoma, stage T1-3 N1-2 M0 there is a statistically significant direct correlation between the number of "points" of macroscopically distinguishable peritumoral lymphangiogenesis and lifespan after a radical surgical treatment. Lymphatic vessels in the process of peritumoral lymphangiogenesis are the indicator of compensatory-protective resources condition.

Prevalence of PD-L1 expression is associated with EMAST, density of peritumoral T-cells and recurrence-free survival in operable non-metastatic colorectal cancer

IntroductionMicrosatellite instability (MSI) predict response to anti-PD1 immunotherapy in colorectal cancer (CRC). CRCs with MSI have higher infiltration of immune cells related to a better survival. Elevated Microsatellite Alterations at Tetranucleotides (EMAST) is a form of MSI but its association with PD-L1 expression and immune-cell infiltration is not known.MethodsA consecutive, observational cohort of patients undergoing surgery for CRC. EMAST and clinicopathological characteristics were investigated against PD-L1, as well as CD3 and CD8 expression in the invasive margin or tumour centre (Immunoscore). Difference in survival between groups was assessed by log rank test.ResultsA total of 149 stage I–III CRCs patients, with a median follow up of 60.1 months. Patients with PD-L1+ tumours (7%) were older (median 79 vs 71 years, p = 0.045) and had EMAST+ cancers (OR 10.7, 95% CI 2.2–51.4, p = 0.001). Recurrence-free survival was longer in cancers with PD-L1+ immune cells (HR 0.35, 95% CI 0.16–0.76, p = 0.008, independent of EMAST) and high Immunoscore (HR 0.10, 95% CI 0.01–0.72, p = 0.022). Patients expressing PD-L1 in immune cells had longer disease-specific survival (HR 0.28, 95% CI 0.10–0.77, p = 0.014).ConclusionsHigher Immunoscore (CD3/CD8 cells) and expression of tumour PD-L1 is found in CRCs with EMAST. Lymphocytic infiltrate and peritumoral PD-L1 expression have prognostic value in CRC.