Interaction Networks Converging on Immunosuppressive Roles of Granzyme B: Special Niches Within the Tumor Microenvironment

Wang, Weinan; Zou, Rui; Qiu, Ye; Liu, Jishuang; Yu, Xin; He, Tianzhu; Qiu, Zhidong

doi:10.3389/fimmu.2021.670324

Cited by 12 publications

(14 citation statements)

References 149 publications

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“…The proinflammatory protease MMP7 is associated with breakdown of the extracellular matrix, wound healing and tumor metastasis ( 42 ). Although granzyme B is known chiefly as an effector molecule which, in conjunction with perforin, mediates T-cell and NK-cell cytotoxic effector responses, soluble granzyme B is also associated with the immunosuppressive tumor niche ( 43 ) and is secreted directly by breast ( 44 ) and urothelial tumor cells, where it’s presence is associated with tumor EMT and invasion ( 45 , 46 ). B7-H6 is associated with tumor progression and metastasis in ovarian ( 47 ) and breast ( 48 ) cancer, but when expressed by tumor, also may promote recognition by NK cells ( 49 ).…”

Section: Discussionmentioning

confidence: 99%

A maladaptive pleural environment suppresses preexisting anti-tumor activity of pleural infiltrating T cells

et al. 2023

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IntroductionTreatment options for patients with malignant pleural effusions (MPE) are limited due, at least in part, to the unique environment of the pleural space, which drives an aggressive tumor state and governs the behavior of infiltrating immune cells. Modulation of the pleural environment may be a necessary step toward the development of effective treatments. We examine immune checkpoint molecule (ICM) expression on pleural T cells, the secretomes of pleural fluid, pleural infiltrating T cells (PIT), and ability to activate PIT ex vivo.MethodsICM expression was determined on freshly drained and in vitro activated PIT from breast, lung and renal cell cancer. Secretomics (63 analytes) of activated PIT, primary tumor cultures and MPE fluid was determined using Luminex technology. Complementary digital spatial proteomic profiling (42 analytes) of CD45+ MPE cells was done using the Nanostring GeoMx platform. Cytolytic activity was measured against autologous tumor targets.ResultsICM expression was low on freshy isolated PIT; regulatory T cells (T-reg) were not detectable by GeoMx. In vitro activated PIT coexpressed PD-1, LAG-3 and TIGIT but were highly cytotoxic against autologous tumor and uniquely secreted cytokines and chemokines in the > 100 pM range. These included CCL4, CCL3, granzyme B, IL-13, TNFα, IL-2 IFNγ, GM-CSF, and perforin. Activated PIT also secreted high levels of IL-6, IL-8 and sIL-6Rα, which contribute to polarization of the pleural environment toward wound healing and the epithelial to mesenchymal transition. Addition of IL-6Rα antagonist to cultures reversed tumor EMT but did not alter PIT activation, cytokine secretion or cytotoxicity.DiscussionDespite the negative environment, immune effector cells are plentiful, persist in MPE in a quiescent state, and are easily activated and expanded in culture. Low expression of ICM on native PIT may explain reported lack of responsiveness to immune checkpoint blockade. The potent cytotoxic activity of activated PIT and a proof-of-concept clinical scale GMP-expansion experiment support their promise as a cellular therapeutic. We expect that a successful approach will require combining cellular therapy with pleural conditioning using immune checkpoint blockers together with inhibitors of upstream master cytokines such as the IL-6/IL-6R axis.

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Section: Discussionmentioning

confidence: 99%

A maladaptive pleural environment suppresses preexisting anti-tumor activity of pleural infiltrating T cells

et al. 2023

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“…The present results indicating that a low amount of GrB+ lymphocytes with respect to tumor-infiltrating CTLs associates with better RFS may be an indication of an immunosuppressive role for GrB in CM. Indeed, GrB is also expressed by a variety of immunosuppressive cells, such as Bregs and Tregs [ 18 ]. For example, GrB+ B cells have been shown to inhibit the proliferation of T cells by degrading the T cell receptor zeta chain, which is a substrate for GrB, in a GrB-dependent manner [ 27 ].…”

Section: Discussionmentioning

confidence: 99%

“…Thus, GrB has been used as a marker for anti-tumor cytolytic activity in, for example, colorectal cancer, where it associates positively with increased survival times [ 16 , 17 ]. However, GrB is also expressed by immunosuppressive cells, such as FoxP3+ Tregs and B regulatory cells (Bregs), to mediate their immunosuppressive effects [ 18 ]. In CM, GrB expression has not been studied extensively.…”

Section: Introductionmentioning

confidence: 99%

“…This study focused on CD8+ and GrB+ lymphocytes, because CD8 is a marker for all cytotoxic T cells and this marker does not correlate with lymphocyte activity. In contrast, GrB is produced by activated lymphocytes, which can represent immunosuppressive Tregs or regulatory B cells (Bregs), or cytotoxic CD8+ T cells and natural killer cells [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

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The Association of CD8+ Cytotoxic T Cells and Granzyme B+ Lymphocytes with Immunosuppressive Factors, Tumor Stage and Prognosis in Cutaneous Melanoma

et al. 2022

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The immunosuppressive tumor microenvironment (TME) consists of suppressive cells producing a variety of immunomodulatory proteins, such as programmed death ligand 1 (PD-L1) and indoleamine-2,3-dioxygenase (IDO). Although granzyme B (GrB) is known to convey the cytolytic activities of CD8+ cytotoxic lymphocytes, it is also expressed by other cells, such as regulatory T and B cells, for immunosuppressive purposes. The role of GrB+ lymphocytes in melanoma has not been examined extensively. In this study, benign, premalignant, and malignant melanocytic tumors were stained immunohistochemically for CD8 and GrB. PD-L1 was also stained from malignant samples that had accompanying clinicopathological data. The association of CD8+ and GrB+ lymphocytes with PD-L1 expression, tumor stage, prognosis, and previously analyzed immunosuppressive factors were evaluated. Our aim was to obtain a more comprehensive perception of the immunosuppressive TME in melanoma. The results show that both CD8+ and GrB+ lymphocytes were more abundant in pT4 compared to pT1 melanomas, and in lymph node metastases compared to primary melanomas. Surprisingly, a low GrB/CD8 ratio was associated with better recurrence-free survival in primary melanomas, which indicates that GrB+ lymphocytes might represent activated immunosuppressive lymphocytes rather than cytotoxic T cells. In the present study, CD8+ lymphocytes associated positively with both tumor and stromal immune cell PD-L1 and IDO expression. In addition, PD-L1+ tumor and stromal immune cells associated positively with IDO+ stromal immune and melanoma cells. The data suggest that IDO and PD-L1 seem to be key immunosuppressive factors in CD8+ lymphocyte-predominant tumors in CM.

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“…In our studies, this truncated tnFGFR1 appears to maintain a stem cell phenotype in hematopoietic stem/precursor cells by regulating expression of genes such as Flt3 and Kit. Clearly, the generation of truncated FGFR1 depends on the expression of Granzyme B which is highly upregulated in hematopoietic cell lineages [36]. The SCLL model we describe here, therefore, offers an ideal opportunity to study the effects of truncated FGFR1-directed gene expression promoting malignant progression in HSCs.…”

Section: Discussionmentioning

confidence: 99%

A truncated derivative of FGFR1 kinase cooperates with FLT3 and KIT to transform hematopoietic stem cells in syndromic and de novo AML

et al. 2022

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Background Myeloid and lymphoid malignancies associated with chimeric FGFR1 kinases are the hallmark of stem cell leukemia and lymphoma syndrome (SCLL). In all cases, FGFR1 kinase is constitutively phosphoactivated as a result of chromosome translocations, which lead to acquisition of dimerization motifs in the chimeric proteins. Recently, we demonstrated that these chimeric kinases could be cleaved by granzyme B to generate a truncated derivative, tnFGFR1, which localized exclusively into the nucleus and was not phosphorylated. Methods Stem cell transduction and transplantation in syngeneic mice was used to assess the transforming ability of tnFGFR1 in bone marrow stem cells, and RPPA and RNA-Seq was used to examine the related signaling pathways and regulated target genes. Results For the first time, we show that this non-classical truncated form of FGFR1 can independently lead to oncogenic transformation of hematopoietic stem cells in an animal model in vivo. These leukemia cells show a mixed immunophenotype with a B-cell B220 + Igm- profile in the majority of cells and Kit+ in virtually all cells, suggesting a stem cell disease. tnFGFR1, however, does not activate classic FGFR1 downstream signaling pathways but induces a distinct profile of altered gene expression with significant upregulation of transmembrane signaling receptors including FLT3 and KIT. We further show that de novo human AML also express tnFGFR1 which correlates with upregulation of FLT3 and KIT as in mouse leukemia cells. ChIP analysis demonstrates tnFGFR1 occupancy at the Flt3 and Kit promoters, suggesting a direct transcriptional regulation. Cells transformed with tnFGFR1 are insensitive to FGFR1 inhibitors but treatment of these cells with the Quizartinib (AC220) FLT3 inhibitor, suppresses in vitro growth and development of leukemia in vivo. Combined treatment with FGFR1 and FLT3 inhibitors provides increased survival compared to FGFR1 inhibition alone. Conclusions This study demonstrates a novel model for transformation of hematopoietic stem cells by chimeric FGFR1 kinases with the combined effects of direct protein activation by the full-length kinases and transcriptional regulation by the truncated nuclear tnFGFR1 derivative, which is associated with GZMB expression levels. Genes significantly upregulated by tnFGFR1 include Flt3 and Kit which promote a leukemia stem cell phenotype. In human AML, tnFGFR1 activation leads to increased FLT3 and KIT expression, and higher FLT3 and GZMB expression levels are associated with an inferior prognosis. These observations provide insights into the relative therapeutic value of targeting FGFR1 and FLT3 in treating AML with this characteristic gene expression profile.

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Interaction Networks Converging on Immunosuppressive Roles of Granzyme B: Special Niches Within the Tumor Microenvironment

Cited by 12 publications

References 149 publications

A maladaptive pleural environment suppresses preexisting anti-tumor activity of pleural infiltrating T cells

A maladaptive pleural environment suppresses preexisting anti-tumor activity of pleural infiltrating T cells

The Association of CD8+ Cytotoxic T Cells and Granzyme B+ Lymphocytes with Immunosuppressive Factors, Tumor Stage and Prognosis in Cutaneous Melanoma

A truncated derivative of FGFR1 kinase cooperates with FLT3 and KIT to transform hematopoietic stem cells in syndromic and de novo AML

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