Modeling T cell temporal response to cancer immunotherapy rationalize development of combinatorial treatments protocols
Ontology highlight
ABSTRACT: Successful immunotherapy relies on triggering complex responses involving T-cells dynamics in tumors and the periphery. Characterizing these responses remains challenging using static human single-cell atlases or mouse models. To address this, we developed a framework for in vivo tracking of tumor-specific CD8+ T cells over time and at single-cell resolution. Our tools facilitate the modeling of gene program dynamics in the tumor microenvironment (TME) and tumor-draining lymph node (tdLN). Using this approach, we characterize two modes of anti-PD1 (aPD1) activity, decoupling induced differentiation of tumor-specific activated precursor cells from cDC1-dependent proliferation and recruitment to the TME. We demonstrate that combining aPD1 with anti-4-1BB agonist enhances the recruitment and proliferation of activated precursors resulting in tumor control. These data suggest that effective response to aPD1 therapy is dependent on sufficient influx of activated precursor CD8+ cells to the TME, and highlight the importance of understanding system-level dynamics in optimizing immunotherapies.
Project description:This work aimed to understand the source of TCF1+ CD8 T cells in tumors. By studying human tumor draining lymph-nodes (TDLNs), we find that CD8 T cells activate in TDLN but fail to acquire the normal effector program. Instead, these CD8 T cells share functional, transcriptional, and epigenetic traits with TCF1+ stem-like cells in the tumor. This suggests that these activated cells are a precursor to the stem-like CD8 T cells in tumors. By using mouse models, we show that tumor-specific CD8 T cells are activated in TDLN, but do not acquire an effector phenotype. These cells then migrate into the tumor where they require additional co-stimulation from antigen presenting cells to acquire the effector program. This is strikingly different from canonical CD8 T cell activation to acute viruses, where the effector program is acquired immediately. This model of T cell differentiation has important implications to improve immunotherapy.
Project description:We previously developed human CAR macrophages (CAR-M) and demonstrated redirection of macrophage anti-tumor function leading to tumor control in immunodeficient xenograft models. Here, we developed clinically relevant fully immunocompetent syngeneic models to evaluate the potential for CAR-M to remodel the tumor microenvironment (TME), induce T cell anti-tumor immunity, and sensitize solid tumors to PD1/PDL1 checkpoint inhibition. In vivo, anti-HER2 CAR-M significantly reduced tumor burden, prolonged survival, remodeled the TME, increased intratumoral T cell and natural killer (NK) cell infiltration, and induced epitope spreading. CAR-M therapy protected against antigen-negative relapse in a T cell dependent fashion, confirming long-term anti-tumor immunity. In HER2+ solid tumors resistant to anti-PD1(aPD1) monotherapy, the combination of CAR-M and aPD1 significantly improved tumor growth control, survival, and remodeling of the TME. These results demonstrate synergy between CAR-M and T cell checkpoint blockade and provide a strategy to enhance response to aPD1 therapy for patients with non-responsive tumors.
Project description:We previously developed human CAR macrophages (CAR-M) and demonstrated redirection of macrophage anti-tumor function leading to tumor control in immunodeficient xenograft models. Here, we developed clinically relevant fully immunocompetent syngeneic models to evaluate the potential for CAR-M to remodel the tumor microenvironment (TME), induce T cell anti-tumor immunity, and sensitize solid tumors to PD1/PDL1 checkpoint inhibition. In vivo, anti-HER2 CAR-M significantly reduced tumor burden, prolonged survival, remodeled the TME, increased intratumoral T cell and natural killer (NK) cell infiltration, and induced epitope spreading. CAR-M therapy protected against antigen-negative relapse in a T cell dependent fashion, confirming long-term anti-tumor immunity. In HER2+ solid tumors resistant to anti-PD1(aPD1) monotherapy, the combination of CAR-M and aPD1 significantly improved tumor growth control, survival, and remodeling of the TME. These results demonstrate synergy between CAR-M and T cell checkpoint blockade and provide a strategy to enhance response to aPD1 therapy for patients with non-responsive tumors.
Project description:In patients with pancreatic ductal adenocarcinoma (PDAC), we show that response to radiation therapy (RT) is characterized by increased IL2R and IL2R expression, decreased ILR2 and exhaustion markers. The bispecific PD-1-targeted IL-2 variant (IL2v) immunocytokine with engineered IL-2 cis-targeted to PD-1 and abolished IL2R binding targets the activation of tumor-antigen specific T cells while rescuing them from Treg suppression. Using aPD1-IL2v in orthotopic PDAC KPC-driven tumor models, we show marked improvement in local and metastatic survival along with profound increase in tumor-infiltrating polyfunctional CD8 T cell subsets with a transcriptionally and metabolically active phenotype, and preferential activation of antigen-specific CD8 T cells. In combination with single dose RT, aPD1-IL2v treatment results in a robust, durable expansion of polyfunctional CD8 T cells, T cell stemness, tumor-specific memory immune response, NK cell activation, and decreased Tregs. These data show that the novel aPD1-IL2v, leads to profound local and distant response in PDAC.
Project description:The recognition of the immune system as a key component of the tumor microenvironment (TME) led to promising therapeutics. Since such therapies benefit only subsets of patients, understanding the activities of immune cells in the TME is required. Eosinophils are an integral part of the TME especially in mucosal tumors. Nonetheless, their role in the TME and the environmental cues that direct their activities are largely unknown, especially in metastasis. We report that breast cancer-driven lung metastasis is characterized by resident and recruited eosinophils. Eosinophil recruitment to the metastatic lung was regulated by G protein coupled receptor signaling but independent of CCR3. Functionally, eosinophils promoted lymphocyte-mediated anti tumor immunity. Transcriptome and proteomic analyses identified the TME rather than intrinsic differences between eosinophil subsets as a key instructing factor directing anti tumorigenic eosinophil activities. Specifically, TNF-a/IFN-g-activated eosinophils facilitated CD4+ and CD8+ T cell infiltration and promoted anti-tumor immunity. Collectively, we identify a mechanism by which the TME entrains eosinophils to adopt anti-tumorigenic properties, which may lead to the development of eosinophil-targeted therapeutics.
Project description:Cervical cancer (CC) is one of the most common malignancy in women worldwide. It is characterized by a natural continuous phenomenon, that is, it is in the initial stage of HPV infection, progresses to intraepithelial neoplasia, and then develops into invasion and metastasis. Determining the complexity of tumor microenvironment (TME) can deepen our understanding of lesion progression and provide novel therapeutic strategies for CC. We performed the single-cell RNA sequencing on the normal cervix, intraepithelial neoplasia, primary tumor and metastatic lymph node tissues to describe the composition, lineage, and functional status of immune cells and mesenchymal cells at different stages of CC progression. A total of 59913 single cells were obtained and divided into 9 cellular clusters, including immune cells (T/NK cells, macrophages, B cells, plasma cells, mast cells and neutrophils) and mesenchymal cells (endothelial cells, smooth muscle cells and fibroblasts). Our results showed that there were distinct cell subpopulations in different stages of CC. High-stage intraepithelial neoplasia (HSIL) tissue exhibited a low, recently activated TME, and it was characterized by high infiltration of tissue-resident CD8 T cell, effector NK cells, Treg, DC1, pDC, and M1-like macrophages. Tumor tissue displayed high enrichment of exhausted CD8 T cells, resident NK cells and M2-like macrophages, suggesting immunosuppressive TME. Metastatic lymph node consisted of naive T cell, central memory T cell, circling NK cells, cytotoxic CD8+ T cells and effector memory CD8 T cells, suggesting an early activated phase of immune response. This study is the first to delineate the transcriptome profile of immune cells during CC progression using single-cell RNA sequencing. Our results indicated that HSIL exhibited a low, recently activated TME, tumor displayed immunosuppressive statue, and metastatic lymph node showed early activated phase of immune response. Our study enhanced the understanding of dynamic change of TME during CC progression and has implications for the development of novel treatments to inhibit the initiation and progression of CC.
Project description:Apoptotic cells are immunosuppressive, creating a potential therapeutic barrier in cancer therapy. In this reportwe investigated responses to apoptotic tumor cell phagocytosis (i.e. efferocytosis) after therapy in the tumor draining lymph node (TDLN). Treatment with cisplatin or the BRAF inhibitor PLX4720 caused a significant increase in accumulation of apoptotic tumor material in the TDLN. We identified the primary phagocyte population responsible for clearing dying tumors in the TDLN was medullary sinus macrophages (MSMs). Tumor cell efferocytosis by MSMs induced an immune suppressive transcriptional program distinct from other macrophage or dendritic cell populations in the TDLN. One of the most differentially induced mRNAs in MSMs wasIl33,coding fora potent immune-regulatory cytokine. Administration of neutralizing anti-IL-33 receptor antibodies or deletion ofIl33in MSMs altered tumor responses to therapy with a significant enhancement of anti-tumor activity compared to controls. Mechanistically, IL-33 activated Treg cells in the TDLN with subsequent suppression of CD8+T cell responses. Importantly, combining IL-33 receptor blockade, BRAF inhibitor, and PD-1 blockade significantly improved tumor regression with enhanced, CD8+T cell dependent immunity. Finally, apoptotic tumor cells induced IL-33 expression in human macrophages, IL-33 expression in sentinel lymph nodes positively corresponded with disease stage and negatively correlated with survival in melanoma and breast cancer. Thus, the data identifies an immune response to therapy that abrogates nascent anti-tumor immunity, providing a previously undescribed functional interaction with broad implications for cancer therapy.
Project description:Apoptotic cells are immunosuppressive, creating a potential therapeutic barrier in cancer therapy. In this reportwe investigated responses to apoptotic tumor cell phagocytosis (i.e. efferocytosis) after therapy in the tumor draining lymph node (TDLN). Treatment with cisplatin or the BRAF inhibitor PLX4720 caused a significant increase in accumulation of apoptotic tumor material in the TDLN. We identified the primary phagocyte population responsible for clearing dying tumors in the TDLN was medullary sinus macrophages (MSMs). Tumor cell efferocytosis by MSMs induced an immune suppressive transcriptional program distinct from other macrophage or dendritic cell populations in the TDLN. One of the most differentially induced mRNAs in MSMs wasIl33,coding fora potent immune-regulatory cytokine. Administration of neutralizing anti-IL-33 receptor antibodies or deletion ofIl33in MSMs altered tumor responses to therapy with a significant enhancement of anti-tumor activity compared to controls. Mechanistically, IL-33 activated Treg cells in the TDLN with subsequent suppression of CD8+T cell responses. Importantly, combining IL-33 receptor blockade, BRAF inhibitor, and PD-1 blockade significantly improved tumor regression with enhanced, CD8+T cell dependent immunity. Finally, apoptotic tumor cells induced IL-33 expression in human macrophages, IL-33 expression in sentinel lymph nodes positively corresponded with disease stage and negatively correlated with survival in melanoma and breast cancer. Thus, the data identifies an immune response to therapy that abrogates nascent anti-tumor immunity, providing a previously undescribed functional interaction with broad implications for cancer therapy.
Project description:We report the transcriptional profiles of tumor-specific CD8+ T cells isolated from tumor draining lymph nodes (TDLN) and tumor following tumor vaccination in C57BL/6 mice. In addition, we included wild type (WT) and TGF-b receptor conditional knockout (Tgfbr2-/-) CD8+ T cells.
Project description:Radiotherapy (RT) and anti-PD-L1 synergize to enhance local and distant (abscopal) tumor control. However, clinical results in humans have been variable. With the goal of improving clinical outcomes, we investigated the underlying synergistic mechanism focusing on a TCF-1+ PD-1+ CD8+ stem-like T cell subset in the tumor-draining lymph node (TdLN). We found that RT + anti-PD-L1 induces a novel differentiation program in the TdLN stem-like population which leads to their expansion and differentiation into effector cells within the tumor. We also found that optimal synergy between RT + anti-PD-L1 is dependent on the TdLN stem-like T cell population as either blockade of TdLN egress or specific stem-like T cell depletion reduced tumor control. Together, these data demonstrate a multistep stimulation of stem-like T cells following combination therapy which is initiated in the TdLN and completed in the tumor.