Prognostic Values of B7-H3, B7-H4, and HHLA2 Expression in Human Pancreatic Cancer Tissues Based on mIHC and Spatial Distribution Analysis

Abstract Background: Pancreatic cancer (PC) is one of the most malignant cancers and its 5-year survival rate remains poor. Although immunotherapy has achieved certain therapeutic efficacy in some clinical trials, such treatment still shows low responsiveness and overall remission rate. Therefore, it is urgently necessary to dissect the tumor microenvironment and optimize the immunotherapeutic strategies against this malignancy.Methods: Using the multi-color immunohistochemistry, we investigated the expressions of B7-H3, B7-H4, HHLA2, CD8, and CD68 in 63 cases of PC tissues with the tissue microarray. Moreover, we analyzed immunolocalization features, prognostic values of this immune contexture, and clinical associations.Results: The expressions of B7-H3, B7-H4, and HHLA2 could be detected in cytokeratin (CK)+ tumor cells, CD68+macrophages, and even stromal cells. Higher expression of B7-H3 in tumor cells could predict a better survival of the PC patients. A positive correlation was found between the expressions of B7-H3 and HHLA2 in tumor cells, while there was a negative correlation between the expressions of B7-H4 and HHLA2 in tumor cells. A positive correlation was found between the expressions of B7-H3 and B7-H4 or HHLA2 in tumor-associated macrophages (TAMs), but not B7-H4 and HHLA2. Tumor-infiltrating CD8+T cells in combination with CD68+TAMs could serve as an important predictor for the postoperative prognosis of PC patients. Higher expression of B7-H3, or HHLA2 in CD68+TAMs could serve as an important predictor for poorer prognosis of PC patients. Patients with B7-H3lowB7-H4low, B7-H3lowHHLA2low, or B7-H4lowHHLA2low on CD68+TAMs could have a better postoperative prognosis compared with the other sub-populations in the combinational analysis.Conclusions: Taken together, our study indicated variable expressions and prognostic values of B7-H3, B7-H4, and HHLA2, in human PC tissues, and distinctively demonstrated that these immunosuppressive co-stimulators expressed by CD68+TAMs could be used as important bio-markers for the prognostic prediction of PC patients. Moreover, these results supported that the evaluation of these markers could be used as essential candidate targets for immunotherapy against PC.

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Patterns and Relevance of Langerhans Islet Invasion in Pancreatic Cancer

Cancers ◽

10.3390/cancers13020249 ◽

2021 ◽

Vol 13 (2) ◽

pp. 249

Author(s):

Ruediger Goess ◽

Ayse Ceren Mutgan ◽

Umut Çalışan ◽

Yusuf Ceyhun Erdoğan ◽

Lei Ren ◽

...

Keyword(s):

Diabetes Mellitus ◽

Pancreatic Cancer ◽

Cancer Cells ◽

Tumor Cells ◽

Islet Cells ◽

Human Pancreatic Cancer ◽

Type I ◽

Type Iv ◽

Pancreatic Cancer Cells

Background: Pancreatic cancer‐associated diabetes mellitus (PC‐DM) is present in most patients with pancreatic cancer, but its pathogenesis remains poorly understood. Therefore, we aimed to characterize tumor infiltration in Langerhans islets in pancreatic cancer and determine its clinical relevance. Methods: Langerhans islet invasion was systematically analyzed in 68 patientswith pancreatic ductal adenocarcinoma (PDAC) using histopathological examination and 3D in vitro migration assays were performed to assess chemoattraction of pancreatic cancer cells to isletcells. Results: Langerhans islet invasion was present in all patients. We found four different patterns of islet invasion: (Type I) peri‐insular invasion with tumor cells directly touching the boundary, but not penetrating the islet; (Type II) endo‐insular invasion with tumor cells inside the round islet; (Type III) distorted islet structure with complete loss of the round islet morphology; and (Type IV)adjacent cancer and islet cells with solitary islet cells encountered adjacent to cancer cells. Pancreatic cancer cells did not exhibit any chemoattraction to islet cells in 3D assays in vitro. Further, there was no clinical correlation of islet invasion using the novel Islet Invasion Severity Score (IISS), which includes all invasion patterns with the occurrence of diabetes mellitus. However, Type IV islet invasion was related to worsened overall survival in our cohort. Conclusions: We systematically analyzed, for the first time, islet invasion in human pancreatic cancer. Four different main patterns of islet invasion were identified. Diabetes mellitus was not related to islet invasion. However, moreresearch on this prevailing feature of pancreatic cancer is needed to better understand underlying principles.

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M1986 Newcastle Disease Virus Lasota Strain Kills Human Pancreatic Cancer Tumor Cells In Vitro With High Selectivity

Gastroenterology ◽

10.1016/s0016-5085(10)62095-1 ◽

2010 ◽

Vol 138 (5) ◽

pp. S-453 ◽

Cited By ~ 2

Author(s):

Robert J. Walter ◽

Bashar M. Attar ◽

Sooraj TK Lakshminarayan ◽

Asad Rafiq ◽

Megan Delimata

Keyword(s):

Pancreatic Cancer ◽

Newcastle Disease Virus ◽

Tumor Cells ◽

Disease Virus ◽

Newcastle Disease ◽

High Selectivity ◽

Human Pancreatic Cancer ◽

Cancer Tumor

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Growth Hormone Receptor Inhibition Sensitizes Human Pancreatic Cancer to Chemotherapy Treatments

Journal of the Endocrine Society ◽

10.1210/jendso/bvab048.2086 ◽

2021 ◽

Vol 5 (Supplement_1) ◽

pp. A1019-A1020

Author(s):

Reetobrata Basu ◽

John Joseph Kopchick ◽

Silvana Duran Ortiz ◽

Yanrong Qian ◽

Prateek Kulkarni

Keyword(s):

Pancreatic Cancer ◽

Growth Hormone ◽

Cancer Cells ◽

Tumor Cells ◽

Hormone Receptor ◽

Growth Hormone Receptor ◽

Human Pancreatic Cancer ◽

Drug Efflux ◽

Pancreatic Cancer Cells

Abstract Human growth hormone (GH) and its cognate growth hormone receptor (GHR) have been established to have a distinct role in promoting the progression of several types of human cancers. We had earlier described a newfound role of the GH-GHR axis in driving chemoresistance in melanoma by upregulating drug efflux by ABC multidrug transporter expression and a phenotype switch by induction of epithelial-to-mesenchymal transition (EMT). Here we present an in-depth analysis of this role of GH-GHR in the highly therapy resistant human pancreatic cancer which has a 5-year survival rate of only 10% in 2020. Using human and mouse pancreatic cancer cells and RNA and protein expression analyses by RT-qPCR, ELISA, and western-blot, we identified that (i) GH upregulates specific ABC-transporter expressions in a drug-context specific manner, (ii) GH upregulates EMT transcription factors, (iii) GH activates specific oncogenic signaling pathways, and (iii) GH action increases cytochrome P450 members involved in hepatic drug metabolism. The GH antagonist, Pegvisomant, significantly inhibited these effects. Additionally, we confirmed the effects of these molecular changes by specific assays. For example, GH increases basement membrane invasion, viability of circulating tumor cells, and drug efflux; while inhibition of GHR by pegvisomant in pancreatic cancer cells reversed this aggressive tumor phenotype and sensitized the tumor cells to chemotherapy. Cell viability assays confirmed a decreased IC50 of gemcitabine, doxorubicin, and erlotinib in pancreatic cancer cells treated with pegvisomant and an increase in IC50 cells treated with GH. We further verified our results using in silico analyses of TCGA datasets for pancreatic cancer - which provided robust confirmation of our experimental findings. Presently we are validating our observation in nude mice with human pancreatic cancer cell xenografts. In conclusion, our in vitro results confirm that GHR antagonism can drastically sensitize human pancreatic cancer cells by blocking mechanisms of drug resistance, thus providing a valuable window for improved efficacy of available chemo- and targeted therapy.

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Silencing of caveolin-1 in fibroblasts as opposed to epithelial tumor cells results in increased tumor growth rate and chemoresistance in a human pancreatic cancer model

International Journal of Oncology ◽

10.3892/ijo.2018.4640 ◽

2018 ◽

Cited By ~ 3

Author(s):

Konstantinos Kamposioras ◽

Chrysiida Tsimplouli ◽

Caroline Verbeke ◽

Alan Anthoney ◽

Argyro Daoukopoulou ◽

...

Keyword(s):

Pancreatic Cancer ◽

Growth Rate ◽

Tumor Growth ◽

Tumor Cells ◽

Human Pancreatic Cancer ◽

Tumor Growth Rate ◽

Cancer Model ◽

Epithelial Tumor ◽

Caveolin 1 ◽

Epithelial Tumor Cells

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A murine model for human pancreatic cancer: Orthotopic injection of syngeneic pancreatic tumor cells

Gastroenterology ◽

10.1016/s0016-5085(98)82647-4 ◽

1998 ◽

Vol 114 ◽

pp. A647

Author(s):

Keita Morikane ◽

Richard M. Tempero ◽

Michael A. Hollingsworth

Keyword(s):

Pancreatic Cancer ◽

Tumor Cells ◽

Murine Model ◽

Pancreatic Tumor ◽

Human Pancreatic Cancer

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Human Pancreatic Cancer Tumors Are Nutrient Poor and Tumor Cells Actively Scavenge Extracellular Protein

Cancer Research ◽

10.1158/0008-5472.can-14-2211 ◽

2015 ◽

Vol 75 (3) ◽

pp. 544-553 ◽

Cited By ~ 326

Author(s):

Jurre J. Kamphorst ◽

Michel Nofal ◽

Cosimo Commisso ◽

Sean R. Hackett ◽

Wenyun Lu ◽

...

Keyword(s):

Pancreatic Cancer ◽

Tumor Cells ◽

Extracellular Protein ◽

Human Pancreatic Cancer

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WDR5-H3K4me3 epigenetic axis regulates OPN expression to compensate PD-L1 function to promote pancreatic cancer immune escape

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2021-002624 ◽

2021 ◽

Vol 9 (7) ◽

pp. e002624

Author(s):

Chunwan Lu ◽

Zhuoqi Liu ◽

John D Klement ◽

Dafeng Yang ◽

Alyssa D Merting ◽

...

Keyword(s):

Pancreatic Cancer ◽

Tumor Growth ◽

Tumor Cells ◽

Protein Level ◽

Pancreatic Tumor ◽

Immune Escape ◽

Epigenetic Mechanism ◽

Human Pancreatic Cancer ◽

Pancreatic Tumors ◽

Genome Wide

BackgroundDespite PD-L1 (Programmed death receptor ligand-1) expression on tumor cells and cytotoxic T lymphocytes tumor infiltration in the tumor microenvironment, human pancreatic cancer stands out as one of the human cancers that does not respond to immune checkpoint inhibitor (ICI) immunotherapy. Epigenome dysregulation has emerged as a major mechanism in T cell exhaustion and non-response to ICI immunotherapy, we, therefore, aimed at testing the hypothesis that an epigenetic mechanism compensates PD-L1 function to render pancreatic cancer non-response to ICI immunotherapy.MethodsTwo orthotopic pancreatic tumor mouse models were used for chromatin immunoprecipitation-Seq and RNA-Seq to identify genome-wide dysregulation of H3K4me3 and gene expression. Human pancreatic tumor and serum were analyzed for osteopontin (OPN) protein level and for correlation with patient prognosis. OPN and PD-L1 cellular location were determined in the tumors using flow cytometry. The function of WDR5-H3K4me3 axis in OPN expression were determined by Western blotting. The function of H3K4me3-OPN axis in pancreatic cancer immune escape and response to ICI immunotherapy was determined in an orthotopic pancreatic tumor mouse model.ResultsMouse pancreatic tumors have a genome-wide increase in H3K4me3 deposition as compared with normal pancreas. OPN and its receptor CD44 were identified being upregulated in pancreatic tumors by their promoter H3K4me3 deposition. OPN protein is increased in both tumor cells and tumor-infiltrating immune cells in human pancreatic carcinoma and is inversely correlated with pancreatic cancer patient survival. OPN is primarily expressed in tumor cells and monocytic myeloid-derived suppressor cells (M-MDSCs), whereas PD-L1 is expressed in tumor cells, M-MDSCs, polymorphonuclear MDSCs and tumor-associated macrophages. WDR5 is essential for H3K4me3-specific histone methyltransferase activity that regulates OPN expression in tumor cells and MDSCs. Inhibition of WDR5 significantly decreased OPN protein level. Inhibition of WDR5 or knocking out of OPN suppressed orthotopic mouse pancreatic tumor growth. Inhibition of WDR5 also significantly increased efficacy of anti-PD-1 immunotherapy in suppression of mouse pancreatic tumor growth in vivo.ConclusionsOPN compensates PD-L1 function to promote pancreatic cancer immune escape. Pharmacological inhibition of the WDR5-H3K4me3 epigenetic axis is effective in suppressing pancreatic tumor immune escape and in improving efficacy of anti-PD-1 immunotherapy in pancreatic cancer.

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Oxygen-Carrying Polymer Nanoconstructs for Radiodynamic Therapy of Deep Hypoxic Malignant Tumors

Biomedicines ◽

10.3390/biomedicines9030322 ◽

2021 ◽

Vol 9 (3) ◽

pp. 322

Author(s):

Sandhya Clement ◽

Anna Guller ◽

Saabah B. Mahbub ◽

Ewa M. Goldys

Keyword(s):

Pancreatic Cancer ◽

Tumor Cells ◽

Cancer Metastasis ◽

Malignant Tumors ◽

Tissue Level ◽

Drug Formulation ◽

Human Pancreatic Cancer ◽

X Rays ◽

Low Oxygen ◽

Pancreatic Cancer Cells

Radiodynamic therapy (RDT) is an emerging non-invasive anti-cancer treatment based on the generation of the reactive oxygen species (ROS) at the lesion site following the interaction between X-rays and a photosensitizer drug (PS). The broader application of RDT is impeded by the tumor-associated hypoxia that results in low availability of oxygen for the generation of sufficient amounts of ROS. Herein, a novel nanoparticle drug formulation for RDT, which addresses the problem of low oxygen availability, is reported. It consists of poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) co-loaded with a PS drug verteporfin (VP), and the clinically approved oxygen-carrying molecule, perfluorooctylbromide (PFOB). When triggered by X-rays (4 Gy), under both normoxic and hypoxic conditions, PLGA–VP–PFOB nanoconstructs (NCs) induced a significant increase of the ROS production compared with matching PLGA–VP nanoparticles. The RDT with NCs effectively killed ~60% of human pancreatic cancer cells in monolayer cultures, and almost completely suppressed the outgrowth of tumor cells in 2-weeks clonogenic assay. In a 3D engineered model of pancreatic cancer metastasis to the liver, RDT with NCs destroyed ~35% of tumor cells, demonstrating an exceptional efficiency at a tissue level. These results show that PLGA–VP–PFOB is a promising agent for RDT of deep-seated hypoxic tumors.

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