53 Unique insights into PDAC development revealed by both InSituPlex® and imaging mass cytometry

BackgroundPancreatic cancer remains a deadly disease due to difficulties hindering its early diagnosis, giving way to metastasis of the tumor and resulting in poor prognosis. While there are many neoplasms of the pancreas, pancreatic invasive ductal adenocarcinoma (PDAC) is the most common and treatment options are few, with poor overall survival. Aggressive surgeries such as the Whipple procedure coupled to systemic chemotherapy is one of the few treatment options. Recently, several publications have demonstrated improved outcomes with the inclusion of immunotherapy to cytotoxic drug combinations in some patients, however optimally selecting patients as candidates for immunotherapy-chemotherapy combinations remains a critical challenge. The complexities of the tumor microenvironment have been implicated in the failure of chemotherapy, radiation therapy, and immunotherapy. The tumor microenvironment of PDAC is especially rich with multiple interactions between pancreatic epithelial/cancer cells, stromal cells, immune cells and the extracellular matrix (ECM). PDACs are characterized by a complex ECM of desmoplastic reaction consisting of an extensive and dense fibrotic stroma that surrounds and infiltrates clusters of malignant epithelial cells, together with the loss of basement membrane integrity and an abnormal vasculature.MethodsIn the present study we demonstrate a tissue phenotyping workflow combining three complementary methods that can unravel novel insights in the complex tumor microenvironment. This novel translational workflow delivers tissue morphology information, spatial phenotyping of immune cell population on whole slides, and high dimensional imaging in selected regions of interest (ROI), by combining H&E, multiplex immunofluorescence (mIF), and Imaging Mass Cytometry (IMC™).ResultsThe use of the InSituPlex® UltiMapper® I/O PD-L1 kit enabled the streamlined combination and alignment of H&E and mIF data, leading to the strategic selection of relevant ROIs, while utility of IMC technology enabled downstream imaging of 35 protein markers associated with the ECM in the selected ROIs to provide a deeper understanding of the tumor microenvironment.ConclusionsThe incorporation of advanced multiplex imaging platforms such as mIF and IMC with routine H&E workflow in tumor biology can deliver some of the much-needed insight into tumor morphology, cellular composition, cellular functions, and cell-cell interactions and paves the way for potentially improved clinical prognosis and efficacy prediction in patients with cancer.

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Suppression of tumor-associated neutrophils by lorlatinib attenuates pancreatic cancer growth and improves treatment with immune checkpoint blockade

Nature Communications ◽

10.1038/s41467-021-23731-7 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Sebastian R. Nielsen ◽

Jan E. Strøbech ◽

Edward R. Horton ◽

Rene Jackstadt ◽

Anu Laitala ◽

...

Keyword(s):

Cd8 T Cells ◽

Immune Cell ◽

Immune Checkpoint Blockade ◽

Ductal Adenocarcinoma ◽

Cancer Growth ◽

Cancer Therapies ◽

Cell Type ◽

Clinical Prognosis ◽

Tumor Associated Neutrophils ◽

Poor Clinical Prognosis

AbstractPancreatic ductal adenocarcinoma (PDAC) patients have a 5-year survival rate of only 8% largely due to late diagnosis and insufficient therapeutic options. Neutrophils are among the most abundant immune cell type within the PDAC tumor microenvironment (TME), and are associated with a poor clinical prognosis. However, despite recent advances in understanding neutrophil biology in cancer, therapies targeting tumor-associated neutrophils are lacking. Here, we demonstrate, using pre-clinical mouse models of PDAC, that lorlatinib attenuates PDAC progression by suppressing neutrophil development and mobilization, and by modulating tumor-promoting neutrophil functions within the TME. When combined, lorlatinib also improves the response to anti-PD-1 blockade resulting in more activated CD8 + T cells in PDAC tumors. In summary, this study identifies an effect of lorlatinib in modulating tumor-associated neutrophils, and demonstrates the potential of lorlatinib to treat PDAC.

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Patient-derived micro-organospheres recapitulate tumor microenvironment and heterogeneity for precision oncology.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.3076 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. 3076-3076

Author(s):

Shengli Ding ◽

Zhaohui Wang ◽

Marcos Negrete Obando ◽

Grecia rivera Palomino ◽

Tomer Rotstein ◽

...

Keyword(s):

Drug Discovery ◽

Tumor Microenvironment ◽

Single Cell ◽

Stromal Cells ◽

Mononuclear Cells ◽

Immune Cell ◽

Tumor Biology ◽

Cell Types ◽

Precision Oncology ◽

Original Tumor

3076 Background: Preclinical models that can recapitulate patients’ intra-tumoral heterogeneity and microenvironment are crucial for tumor biology research and drug discovery. In particular, the ability to retain immune and other stromal cells in the microenvironment is vital for the development of immuno-oncology assays. However, current patient-derived organoid (PDO) models are largely devoid of immune components. Methods: We first developed an automated microfluidic and membrane platform that can generate tens of thousands of micro-organospheres from resected or biopsied clinical tumor specimens within an hour. We next characterized growth rate and drug response of micro-organospheres. Finally, extensive single-cell RNA-seq profiling were performed on both micro-organospheres and original tumor samples from lung, ovarian, kidney, and breast cancer patients. Results: Micro-organospheres derived from clinical tumor samples preserved all original tumor and stromal cells, including fibroblasts and all immune cell types. Single-cell analysis revealed that unsupervised clustering of tumor and non-tumor cells were identical between original tumors and the derived micro-organospheres. Quantification showed similar cell composition and percentages for all cell types and also preserved functional intra-tumoral heterogeneity.. An automated, end-to-end, high-throughput drug screening pipeline demonstrated that matched peripheral blood mononuclear cells (PBMCs) from the same patient added to micro-organospheres can be used to assess the efficacy of immunotherapy moieties. Conclusions: Micro-organospheres are a rapid and scalable platform to preserve patient tumor microenvironment and heterogeneity. This platform will be useful for precision oncology, drug discovery, and immunotherapy development. Funding sources: NIH U01 CA217514, U01 CA214300, Duke Woo Center for Big Data and Precision Health

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LncRNAs Target Ferroptosis-Related Genes and Impair Activation of CD4+ T Cell in Gastric Cancer

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.797339 ◽

2021 ◽

Vol 9 ◽

Author(s):

Fuwen Yao ◽

Yongqiang Zhan ◽

Zuhui Pu ◽

Ying Lu ◽

Jiao Chen ◽

...

Keyword(s):

Gastric Cancer ◽

T Cell ◽

Tumor Microenvironment ◽

T Cell Activation ◽

Cell Activation ◽

Immune Cell ◽

Cell Activity ◽

Cell Receptor ◽

Cell Infiltration ◽

Poor Overall Survival

Gastric cancer (GC) is a malignant disease of the digestive tract and a life-threatening disease worldwide. Ferroptosis, an iron-dependent cell death caused by lipid peroxidation, is reported to be highly correlated with gastric tumorigenesis and immune cell activity. However, the underlying relationship between ferroptosis and the tumor microenvironment in GC and potential intervention strategies have not been unveiled. In this study, we profiled the transcriptome and prognosis data of ferroptosis-related genes (FRGs) in GC samples of the TCGA-STAD dataset. The infiltrating immune cells in GC were estimated using the CIBERSORT and XCELL algorithms. We found that the high expression of the hub FRGs (MYB, PSAT1, TP53, and LONP1) was positively correlated with poor overall survival in GC patients. The results were validated in an external GC cohort (GSE62254). Further immune cell infiltration analysis revealed that CD4+ T cells were the major infiltrated cells in the tumor microenvironment of GC. Moreover, the hub FRGs were significantly positively correlated with activated CD4+ T cell infiltration, especially Th cells. The gene features in the high-FRG score group were enriched in cell division, DNA repair, protein folding, T cell receptor, Wnt and NIK/NF-kappaB signaling pathways, indicating that the hub FRGs may mediate CD4+ T cell activation by these pathways. In addition, an upstream transcriptional regulation network of the hub FRGs by lncRNAs was also developed. Three lncRNAs (A2M-AS1, C2orf27A, and ZNF667-AS1) were identified to be related to the expression of the hub FRGs. Collectively, these results showed that lncRNA A2M-AS1, C2orf27A, and ZNF667-AS1 may target the hub FRGs and impair CD4+ T cell activation, which finally leads to poor prognosis of GC. Effective interventions for the above lncRNAs and the hub FRGs can help promote CD4+ T cell activation in GC patients and improve the efficacy of immunotherapy. These findings provide a novel idea of GC immunotherapy and hold promise for future clinical application.

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Modeling the Tumor Microenvironment of Ovarian Cancer: The Application of Self-Assembling Biomaterials

Cancers ◽

10.3390/cancers13225745 ◽

2021 ◽

Vol 13 (22) ◽

pp. 5745

Author(s):

Ana Karen Mendoza-Martinez ◽

Daniela Loessner ◽

Alvaro Mata ◽

Helena S. Azevedo

Keyword(s):

Ovarian Cancer ◽

Tumor Microenvironment ◽

Materials Science ◽

Treatment Options ◽

Tumor Biology ◽

3D Models ◽

Gynecologic Malignancies ◽

Cell Behaviors ◽

Self Assembling ◽

Key Driver

Ovarian cancer (OvCa) is one of the leading causes of gynecologic malignancies. Despite treatment with surgery and chemotherapy, OvCa disseminates and recurs frequently, reducing the survival rate for patients. There is an urgent need to develop more effective treatment options for women diagnosed with OvCa. The tumor microenvironment (TME) is a key driver of disease progression, metastasis and resistance to treatment. For this reason, 3D models have been designed to represent this specific niche and allow more realistic cell behaviors compared to conventional 2D approaches. In particular, self-assembling peptides represent a promising biomaterial platform to study tumor biology. They form nanofiber networks that resemble the architecture of the extracellular matrix and can be designed to display mechanical properties and biochemical motifs representative of the TME. In this review, we highlight the properties and benefits of emerging 3D platforms used to model the ovarian TME. We also outline the challenges associated with using these 3D systems and provide suggestions for future studies and developments. We conclude that our understanding of OvCa and advances in materials science will progress the engineering of novel 3D approaches, which will enable the development of more effective therapies.

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Analysis of Tumor Microenvironment Characteristics in Bladder Cancer: Implications for Immune Checkpoint Inhibitor Therapy

Frontiers in Immunology ◽

10.3389/fimmu.2021.672158 ◽

2021 ◽

Vol 12 ◽

Author(s):

Xingyu Chen ◽

Haotian Chen ◽

Dong He ◽

Yaxin Cheng ◽

Yuxing Zhu ◽

...

Keyword(s):

Bladder Cancer ◽

Tumor Microenvironment ◽

Cancer Progression ◽

Immune Checkpoint ◽

Immune Checkpoint Inhibitor ◽

Immune Cell ◽

Cox Regression ◽

Prognostic Biomarker ◽

Checkpoint Inhibitor ◽

Clinical Prognosis

The tumor microenvironment (TME) plays a crucial role in cancer progression and recent evidence has clarified its clinical significance in predicting outcomes and efficacy. However, there are no studies on the systematic analysis of TME characteristics in bladder cancer. In this study, we comprehensively evaluated the TME invasion pattern of bladder cancer in 1,889 patients, defined three different TME phenotypes, and found that different subtypes were associated with the clinical prognosis and pathological characteristics of bladder cancer. We further explored the signaling pathways, cancer-immunity cycle, copy number, and somatic mutation differences among the different subtypes and used the principal component analysis algorithm to calculate the immune cell (IC) score, a tool for comprehensive evaluation of TME. Univariate and multivariate Cox regression analyses showed that ICscore is a reliable and independent prognostic biomarker. In addition, the use of anti-programmed death-ligand (PD-L1) treatment cohort, receiver operating characteristic (ROC) curve, Tumor Immune Dysfunction and Exclusion (TIDE), Subnetwork Mappings in Alignment of Pathways (SubMAP), and other algorithms confirmed that ICscore is a reliable prognostic biomarker for immune checkpoint inhibitor response. Patients with higher ICscore showed a significant therapeutic advantage in immunotherapy. In conclusion, this study improves our understanding of the characteristics of TME infiltration in bladder cancer and provides guidance for more effective personalized immunotherapy strategies.

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Molecular Characterization of the Clinical and Tumor Immune Microenvironment Signature of 5-methylcytosine-Related Regulators in non-small Cell Lung Cancer

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.779367 ◽

2021 ◽

Vol 9 ◽

Author(s):

Taisheng Liu ◽

Liyi Guo ◽

Guihong Liu ◽

Xiaoshan Hu ◽

Xiaoning Li ◽

...

Keyword(s):

Lung Cancer ◽

Tumor Microenvironment ◽

Immune Cell ◽

Cell Infiltration ◽

Immune Cell Infiltration ◽

Check Point ◽

Clinical Prognosis ◽

Tumor Mutation Burden ◽

Mutation Burden ◽

Immune Check Point Inhibitor

Background: DNA methylation is an important epigenetic modification, among which 5-methylcytosine methylation (5mC) is generally associated with tumorigenesis. Nonetheless, the potential roles of 5mC regulators in the tumor microenvironment (TME) remain unclear.Methods: The 5mC modification patterns of 1,374 lung adenocarcinoma samples were analyzed systematically. The correlation between the 5mC modification and tumor microenvironment cell infiltration was further assessed. The 5mCscore was developed to evaluate tumor mutation burden, immune check-point inhibitor response, and the clinical prognosis of individual tumors.Results: Three 5mC modification patterns were established based on the clinical characteristics of 21 5mC regulators. According to the differential expression of 5mC regulators, three distinct 5mC gene cluster were also identified, which showed distinct TME immune cell infiltration patterns and clinical prognoses. The 5mCscore was constructed to evaluate the tumor mutation burden, immune check-point inhibitor response, and prognosis characteristics. We found that patients with a low 5mCscore had significant immune cell infiltration and increased clinical benefit.Conclusion: This study indicated that the 5mC modification is involved in regulating TME infiltration remodeling. Targeting 5mC modification regulators might be a novel strategy to treat lung cancer.

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Immune Evasion Mechanism and AXL

Frontiers in Oncology ◽

10.3389/fonc.2021.756225 ◽

2021 ◽

Vol 11 ◽

Author(s):

Hye-Youn Son ◽

Hwan-Kyu Jeong

Keyword(s):

Tumor Microenvironment ◽

Immune Responses ◽

Signaling Pathway ◽

Immune Evasion ◽

Cell Activation ◽

Immune Cell ◽

Nk Cell ◽

Multiple Cancer ◽

Clinical Prognosis ◽

Programmed Death

Extensive interest in cancer immunotherapy is reported according to the clinical importance of CTLA-4 and (PD-1/PD-L1) [programmed death (PD) and programmed death-ligand (PD-L1)] in immune checkpoint therapies. AXL is a receptor tyrosine kinase expressed in different types of cancer and in relation to resistance against various anticancer therapeutics due to poor clinical prognosis. AXL and its ligand, i.e., growth arrest-specific 6 (GAS6) proteins, are expressed on many cancer cells, and the GAS6/AXL pathway is reported to promote cancer cell proliferation, survival, migration, invasion, angiogenesis, and immune evasion. AXL is an attractive and novel therapeutic target for impairing tumor progression from immune cell contracts in the tumor microenvironment. The GAS6/AXL pathway is also of interest immunologically because it targets fewer antitumor immune responses. In effect, several targeted therapies are selective and nonselective for AXL, which are in preclinical and clinical development in multiple cancer types. Therefore, this review focuses on the role of the GAS6/AXL signaling pathway in triggering the immunosuppressive tumor microenvironment as immune evasion. This includes regulating its composition and activating T-cell exclusion with the immune-suppressive activity of regulatory T cells, which is related to one of the hallmarks of cancer survival. Finally, this article discusses the GAS6/AXL signaling pathway in the context of several immune responses such as NK cell activation, apoptosis, and tumor-specific immunity, especially PD-1/PDL-1 signaling.

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Chemokine-Directed Tumor Microenvironment Modulation in Cancer Immunotherapy

International Journal of Molecular Sciences ◽

10.3390/ijms22189804 ◽

2021 ◽

Vol 22 (18) ◽

pp. 9804

Author(s):

Pedro Bule ◽

Sandra I. Aguiar ◽

Frederico Aires-Da-Silva ◽

Joana N. R. Dias

Keyword(s):

Tumor Microenvironment ◽

Immune Cells ◽

Immune Cell ◽

Vascular Endothelial Cells ◽

Tumor Biology ◽

Large Family ◽

Response To Therapy ◽

Cell Trafficking ◽

Immunotherapy Target ◽

Patient Prognosis

Chemokines are a large family of small chemotactic cytokines that coordinates immune cell trafficking. In cancer, they have a pivotal role in the migration pattern of immune cells into the tumor, thereby shaping the tumor microenvironment immune profile, often towards a pro-tumorigenic state. Furthermore, chemokines can directly target non-immune cells in the tumor microenvironment, including cancer, stromal and vascular endothelial cells. As such, chemokines participate in several cancer development processes such as angiogenesis, metastasis, cancer cell proliferation, stemness and invasiveness, and are therefore key determinants of disease progression, with a strong influence in patient prognosis and response to therapy. Due to their multifaceted role in the tumor immune response and tumor biology, the chemokine network has emerged as a potential immunotherapy target. Under the present review, we provide a general overview of chemokine effects on several tumoral processes, as well as a description of the currently available chemokine-directed therapies, highlighting their potential both as monotherapy or in combination with standard chemotherapy or other immunotherapies. Finally, we discuss the most critical challenges and prospects of developing targeted chemokines as therapeutic options.

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A randomized phase II study of cabiralizumab (cabira) + nivolumab (nivo) ± chemotherapy (chemo) in advanced pancreatic ductal adenocarcinoma (PDAC).

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.4_suppl.tps465 ◽

2019 ◽

Vol 37 (4_suppl) ◽

pp. TPS465-TPS465 ◽

Cited By ~ 9

Author(s):

Andrea Wang-Gillam ◽

Eileen Mary O'Reilly ◽

Johanna C. Bendell ◽

Zev A. Wainberg ◽

Erkut Hasan Borazanci ◽

...

Keyword(s):

T Cell ◽

Immune Modulation ◽

Immune Cell ◽

Treatment Options ◽

Objective Response ◽

Locally Advanced ◽

Cell Activity ◽

Progression Free Survival ◽

Ductal Adenocarcinoma ◽

Open Label

TPS465 Background: Treatment options for PDAC are limited; thus, new therapies that can improve outcomes and extend survival are needed. PDAC is associated with high infiltration by tumor-associated macrophages (TAMs) that inhibit antitumor T-cell activity. Blocking colony-stimulating factor 1 receptor (CSF-1R) signaling—which supports the recruitment, differentiation, and maintenance of immunosupressive macrophages in tumors—may lead to depletion of TAMs and upregulation of T-cell checkpoints. Cabira, a humanized IgG4 monoclonal antibody, binds to CSF-1R and blocks its signaling, a key determinant of TAM activation and survival. By reducing TAMs and promoting a proinflammatory microenvironment, cabira may stimulate T-cell responses, thereby sensitizing PDAC to therapy with nivo (anti‒PD-1). In a phase 1a/b study cabira + nivo was tolerable and showed evidence of on-target tumor immune modulation and durable clinical benefit in heavily pretreated patients (pts) with advanced PDAC (Wainberg et al. J Immunother Cancer. 2017 [abst O42]; Carleton et al. J Clin Oncol. 2018 [abst 3020]). Here we describe a randomized, open-label, phase 2 study evaluating the safety and efficacy of cabira + nivo ± chemo in advanced PDAC. Methods: Pts aged ≥18 y with locally advanced/metastatic PDAC that progressed on/after first-line chemo (gemcitabine [gem] or 5-fluorouracil [5-FU] based) will be enrolled. Pts with active/suspected autoimmune disease, uncontrolled/significant cardiovascular disease, or prior exposure to select immune cell–modulating antibodies are not eligible. Depending on prior chemo received, pts will be randomized to 1 of 4 arms (n≈40 each): cabira + nivo; cabira + nivo + gem/nab-paclitaxel; cabira + nivo + oxaliplatin/5-FU/leucovorin; or investigator’s choice of standard-of-care chemo. Endpoints include median progression-free survival (primary), overall survival rate, objective response rate, median duration of response, pharmacokinetics, and safety. In a completed preliminary safety cohort, 12 pts were treated with cabira + nivo + chemo and monitored for 4 wk; competitive enrollment is open, with 32 pts enrolled. (NCT03336216, NCT02526017) Clinical trial information: NCT03336216.

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