The coronavirus disease 2019 (COVID-19) pandemic poses a current world-wide public health threat. However, little is known about its hallmarks compared to other infectious diseases. Here, we report ...the single-cell transcriptional landscape of longitudinally collected peripheral blood mononuclear cells (PBMCs) in both COVID-19- and influenza A virus (IAV)-infected patients. We observed increase of plasma cells in both COVID-19 and IAV patients and XIAP associated factor 1 (XAF1)-, tumor necrosis factor (TNF)-, and FAS-induced T cell apoptosis in COVID-19 patients. Further analyses revealed distinct signaling pathways activated in COVID-19 (STAT1 and IRF3) versus IAV (STAT3 and NFκB) patients and substantial differences in the expression of key factors. These factors include relatively increase of interleukin (IL)6R and IL6ST expression in COVID-19 patients but similarly increased IL-6 concentrations compared to IAV patients, supporting the clinical observations of increased proinflammatory cytokines in COVID-19 patients. Thus, we provide the landscape of PBMCs and unveil distinct immune response pathways in COVID-19 and IAV patients.
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•We generated a single-cell atlas of PBMCs in both COVID-19 and influenza patients•Plasma cells increase significantly in both COVID-19 and influenza patients•COVID-19 is featured with XAF1-, TNF-, and FAS-induced T cell apoptosis•COVID-19 activates distinct pathway (STAT1/IRF3) versus influenza (STAT3/NFκB)
COVID-19 and influenza are both respiratory infections with cytokine release syndrome. Zhu et al. use single-cell RNA sequencing of longitudinally collected PBMCs in both patients to reveal distinct immune response landscapes of the two diseases and identify virus-specific cell composition and immune response pathways.
Macrophages and neutrophils are the main components of the innate immune system and play important roles in promoting angiogenesis, extracellular matrix remodeling, cancer cell proliferation, and ...metastasis in the tumor microenvironment (TME). They can also be harnessed to mediate cytotoxic tumor killing effects and orchestrate effective anti-tumor immune responses with proper stimulation and modification. Therefore, macrophages and neutrophils have strong potential in cancer immunotherapy. In this review, we briefly outlined the applications of macrophages or neutrophils in adoptive cell therapies, and focused on chimeric antigen receptor (CAR)-engineered macrophages (CAR-Ms) and neutrophils (CAR-Ns). We summarized the construction strategies, the preclinical and clinical studies of CAR-Ms and CAR-Ns. In the end, we briefly discussed the limitations and challenges of CAR-Ms and CAR-Ns, as well as future research directions to extend their applications in treating solid tumors.
Receptor-type tyrosine-protein phosphatase T (PTPRT) is a transmembrane protein that is involved in cell adhesion. We previously found that PTPRT was downregulated in multiple cancer types and the ...mutation of PTPRT was associated with cancer early metastasis. However, the impacts of PTPRT downregulation on tumour proliferation, invasion, and clinical interventions such as immune checkpoint inhibitor (ICI) therapies remained largely unknown.
Gene expression data of non-small cell lung cancer (NSCLC) samples from The Cancer Genome Atlas database were downloaded and used to detect the differential expressed genes between PTPRT-high and PTPRT-low subgroups. Knockdown and overexpress of PTPRT in lung cancer cell lines were performed to explore the function of PTPRT in vitro. Western blot and qRT-PCR were used to evaluate the expression of cell cycle-related genes. CCK-8 assays, wound-healing migration assay, transwell assay, and colony formation assay were performed to determine the functional impacts of PTPRT on cell proliferation, migration, and invasion. KM-plotter was used to explore the significance of selected genes on patient prognosis.
PTPRT was found to be downregulated in tumours and lung cancer cell lines compared to normal samples. Cell cycle-related genes (BIRC5, OIP5, and CDCA3, etc.) were specifically upregulated in PTPRT-low lung adenocarcinoma (LUAD). Modulation of PTPRT expression in LUAD cell lines affected the expression of BIRC5 (survivin) significantly, as well as the proliferation, migration, and invasion of tumour cells. In addition, low PTPRT expression level was correlated with worse prognosis of lung cancer and several other cancer types. Furthermore, PTPRT downregulation was associated with elevated tumour mutation burden and tumour neoantigen burden in lung cancer, indicating the potential influence on tumour immunogenicity.
Our findings uncovered the essential roles of PTPRT in the regulation of proliferation, migration, and invasion of LUAD, and highlighted the clinical significance of PTPRT downregulation in lung cancer.
Cancer patients have been treated with various types of therapies, including conventional strategies like chemo‐, radio‐, and targeted therapy, as well as immunotherapy like checkpoint inhibitors, ...vaccine and cell therapy etc. Among the therapeutic alternatives, T‐cell therapy like CAR‐T (Chimeric Antigen Receptor Engineered T cell) and TCR‐T (T Cell Receptor Engineered T cell), has emerged as the most promising therapeutics due to its impressive clinical efficacy. However, there are many challenges and obstacles, such as immunosuppressive tumor microenvironment, manufacturing complexity, and poor infiltration of engrafted cells, etc still, need to be overcome for further treatment with different forms of cancer. Recently, the antitumor activities of CAR‐T and TCR‐T cells have shown great improvement with the utilization of CRISPR/Cas9 gene editing technology. Thus, the genome editing system could be a powerful genetic tool to use for manipulating T cells and enhancing the efficacy of cell immunotherapy. This review focuses on pros and cons of various gene delivery methods, challenges, and safety issues of CRISPR/Cas9 gene editing application in T‐cell‐based immunotherapy.
This review focuses on pros and cons of various gene delivery methods, challenges and safety issues of CRISPR/Cas9 gene editing application in T‐cell based immunotherapy.
T cell receptor-engineered T cells (TCR-Ts) have emerged as potent cancer immunotherapies. While most research focused on classical cytotoxic CD8
+
T cells, the application of CD4
+
T cells in ...adoptive T cell therapy has gained much interest recently. However, the cytotoxic mechanisms of CD4
+
TCR-Ts have not been fully revealed. In this study, we obtained an MHC class I-restricted MART-1
27-35
-specific TCR sequence based on the single-cell V(D)J sequencing technology, and constructed MART-1
27-35
-specific CD4
+
TCR-Ts and CD8
+
TCR-Ts. The antitumor effects of CD4
+
TCR-Ts were comparable to those of CD8
+
TCR-Ts
in vitro
and
in vivo
. To delineate the killing mechanisms of cytotoxic CD4
+
TCR-Ts, we performed single-cell RNA sequencing and found that classical granule-dependent and independent cytolytic pathways were commonly used in CD4
+
and CD8
+
TCR-Ts, while high expression of
LTA
and various costimulatory receptors were unique features for cytotoxic CD4
+
TCR-Ts. Further signaling pathway analysis revealed that transcription factors Runx3 and Blimp1/Tbx21 were crucial for the development and killing function of cytotoxic CD4
+
T cells. Taken together, we report the antitumor effects and multifaceted killing mechanisms of CD4
+
TCR-Ts, and also indicate that MHC class I-restricted CD4
+
TCR-Ts could serve as potential adoptive T cell therapies.
The preexistence of immune cells in the tumor microenvironment substantiates the efficacy of immunotherapy in cancer patients. Although the complex intratumoral immune heterogeneity has been ...extensively studied in single cell resolution, hi-res spatial investigations are limited. In this study, we performed a spatial transcriptome analysis of 4 colorectal adenocarcinoma specimens and 2 paired distant normal specimens to identify the molecular pattern involved in a discontinuous inflammatory response in pathologically annotated cancer regions. Based on the location of spatially varied gene expression, we unmasked the spatially-varied immune ecosystem and identified the locoregional “warmed-up” immune response in predefined “cold” tumor with substantial infiltration of immune components. This “warmed-up” immune profile was found to be associated with the in-situ copy number variance and the tissue remodeling process. Further, “warmed-up” signature genes indicated improved overall survival in CRC patients obtained from TCGA database.
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Neoantigens derived from somatic mutations in cancer cells can induce antigen‐specific T‐cell immune response for cancer immunotherapy. However, the 3D models for assessing neoepitope immunogenicity ...and efficacy of anti‐tumor T‐cell immune response to neoantigens are less than perfect. Here, a 3D tumor model based on recellularized liver matrix is leveraged with HepG2 cells to investigate T cell cytotoxic reactivity toward hepatocellular carcinoma (HCC) neoantigens. The whole exome sequencing (WES) data of 364 HCC patients in The Cancer Genome Atlas database are collected and 25 highly potential immunogenic neoantigens to human leukocyte antigen (HLA)‐A*02:01 molecule in silico are predicted. Six of the HCC neoantigen candidates are functionally validated with high immunogenicity by measuring cellular interferon‐γ secretion and cytotoxicity during neoantigen‐specific T‐cell immune responses in vitro. Then, the minigene of six functionally identified neoantigen peptides is constructed and the minigene‐modified GFP‐HepG2 cells are generated. Neoantigen‐specific immune response is observed with highly secreted Granzyme B, IFN‐γ, and PD‐1 when targeting the minigene‐modified GFP‐HepG2 cells in the 3D RLM HCC tumor model. Overall, the 3D RLM tumor model provides a novel strategy for preclinical assessment of the efficacy of neoantigen‐specific T cell immune response, which helps develop personalized cancer vaccines and immunotherapy treatments for HCC patients.
A 3D tumor model based on a recellularized liver matrix, which incorporates multiple sets of complex perfusable vasculatures, is developed to provide a unique organ‐structured platform that enables cytotoxic T lymphocytes (CTLs) to infiltrate the tumor, recognize neoantigens, and kill cancer cells, replicating the key steps for the CTLs to execute their functions in vivo.
Background
Cancer‐associated fibroblasts (CAFs) are potential targets for cancer therapy. Due to the heterogeneity of CAFs, the influence of CAF subpopulations on the progression of lung cancer is ...still unclear, which impedes the translational advances in targeting CAFs.
Methods
We performed single‐cell RNA sequencing (scRNA‐seq) on tumour, paired tumour‐adjacent, and normal samples from 16 non‐small cell lung cancer (NSCLC) patients. CAF subpopulations were analyzed after integration with published NSCLC scRNA‐seq data. SpaTial enhanced resolution omics‐sequencing (Stereo‐seq) was applied in tumour and tumour‐adjacent samples from seven NSCLC patients to map the architecture of major cell populations in tumour microenvironment (TME). Immunohistochemistry (IHC) and multiplexed IHC (mIHC) were used to validate marker gene expression and the association of CAFs with immune infiltration in TME.
Results
A subcluster of myofibroblastic CAFs, POSTN+ CAFs, were significantly enriched in advanced tumours and presented gene expression signatures related to extracellular matrix remodeling, tumour invasion pathways and immune suppression. Stereo‐seq and mIHC demonstrated that POSTN+ CAFs were in close localization with SPP1+ macrophages and were associated with the exhausted phenotype and lower infiltration of T cells. POSTN expression or the abundance of POSTN+ CAFs were associated with poor prognosis of NSCLC.
Conclusions
Our study identified a myofibroblastic CAF subpopulation, POSTN+ CAFs, which might associate with SPP1+ macrophages to promote the formation of desmoplastic architecture and participate in immune suppression. Furthermore, we showed that POSTN+ CAFs associated with cancer progression and poor clinical outcomes and may provide new insights on the treatment of NSCLC.
POSTN+ CAFs were enriched in advanced NSCLC tumours and presented gene expression signatures related to extracellular matrix remodeling, tumour invasion and immune suppression.
POSTN+ CAFs were in close localization with SPP1+ macrophages and associated with the exhausted phenotype and lower infiltration of T cells.
POSTN expression and POSTN+ CAFs were prognostic factors for NSCLC.
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