Journal Articles

Development of Inducible CD19-CAR T Cells with a Tet-On System for Controlled Activity and Enhanced Clinical Safety

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The tetracycline regulatory system has been widely used to control the transgene expression. With this powerful tool, it might be possible to effectively control the functional activity of chimeric antigen receptor (CAR) T cells and manage the severe side effects after infusion. In this study, we developed novel inducible CD19CAR (iCAR19) T cells by incorporating a one-vector Tet-on system into the CD19CAR construct. The iCAR19 T cells showed dox-dependent cell proliferation, cytokine production, CAR expression, and strong CD19-specific cytotoxicity. After 48 h of dox induction, the relative CAR expression of induced cells was five times greater than that of uninduced cells. Twenty-four hours after dox removal, CAR expression significantly decreased by more than 60%. In cytotoxicity assays, dox-treated cells induced significantly higher specific lysis against target cells. These results suggested that the activity of iCAR19 T cells was successfully controlled by our Tet-on system, offering an enhanced safety profile while maintaining a robust anti-tumor effect. Besides, all manufacture processes of the lentiviral vectors and the T cells were conducted according to the Good Manufacturing Practice (GMP) standards for subsequent clinical translation.

Author Info: (1) International Research Center for Marine Biosciences at Shanghai Ocean University, Ministry of Science and Technology, Shanghai 201306, China. xingjian.gu@mavs.uta.edu. National Pathogen Collection Center for

Author Info: (1) International Research Center for Marine Biosciences at Shanghai Ocean University, Ministry of Science and Technology, Shanghai 201306, China. xingjian.gu@mavs.uta.edu. National Pathogen Collection Center for Aquatic Animals, Shanghai 201306, China. xingjian.gu@mavs.uta.edu. Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding, Shanghai 201306, China. xingjian.gu@mavs.uta.edu. Shanghai Telebio Biomedical Co., Ltd., Shanghai 201321, China. xingjian.gu@mavs.uta.edu. (2) Shanghai Telebio Biomedical Co., Ltd., Shanghai 201321, China. hdy_telebio@163.com. (3) Shanghai Telebio Biomedical Co., Ltd., Shanghai 201321, China. cx-l824@163.com. (4) Shanghai Telebio Biomedical Co., Ltd., Shanghai 201321, China. vector.telebio@163.com. (5) International Research Center for Marine Biosciences at Shanghai Ocean University, Ministry of Science and Technology, Shanghai 201306, China. ghyang119@163.com. National Pathogen Collection Center for Aquatic Animals, Shanghai 201306, China. ghyang119@163.com. Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding, Shanghai 201306, China. ghyang119@163.com. Shanghai Telebio Biomedical Co., Ltd., Shanghai 201321, China. ghyang119@163.com.

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Inhibition of TAMs improves the response to docetaxel in castration-resistant prostate cancer

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For men with castration-resistant prostate cancer (CRPC), androgen-deprivation therapy (ADT) often becomes ineffective requiring the addition of docetaxel, a proven effective chemotherapy option. Tumor-associated macrophages (TAMs) are known to provide protumorigenic influences that contribute to treatment failure. In this study, we examined the contribution of TAMs to docetaxel treatment. An increased infiltration of macrophages in CRPC tumors was observed after treatment with docetaxel. Prostate cancer cells treated with docetaxel released more macrophage colony-stimulating factor (M-CSF-1 or CSF-1), IL-10 and other factors, which can recruit and modulate circulating monocytes to promote their protumorigenic functions. Inhibition of CSF-1 receptor kinase signaling with a small molecule antagonist (PLX3397) in CRPC models significantly reduces the infiltration of TAMs and their influences. As such, the addition of PLX3397 to docetaxel treatment resulted in a more durable tumor growth suppression than docetaxel alone. This study reveals a rational strategy to abrogate the influences of TAMs and extend the treatment response to docetaxel in CRPC.

Author Info: (1) Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. (2) Department of Urology

Author Info: (1) Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. (2) Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. Department of Paediatric Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, California, USA. (3) Department of Urology, Institute of Urology, West China Hospital of Sichuan University, Chengdu, China. (4) Department of Urology, Institute of Urology, West China Hospital of Sichuan University, Chengdu, China. (5) Department of Urology, Institute of Urology, West China Hospital of Sichuan University, Chengdu, China. (6) Plexxikon Inc., Berkeley, California, USA. (7) Plexxikon Inc., Berkeley, California, USA. (8) Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. (9) Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, California, USA. Department of Urology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, California, USA. Department of Pediatrics, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, California, USA. Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, California, USA. Molecular Biology Institute, University of California at Los Angeles, Los Angeles, California, USA.

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Antifibrotic therapy disrupts stromal barriers and modulates the immune landscape in pancreatic ductal adenocarcinoma

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Pancreatic ductal adenocarcinoma (PDA) remains one of the deadliest forms of cancer, in part, because it is largely refractory to current therapies. The failure of most standard therapies in PDA, as well as promising immune therapies, may be largely ascribed to highly unique and protective stromal microenvironments that present significant biophysical barriers to effective drug delivery, that are immunosuppressive, and that can limit the distribution and function of anti-tumor immune cells. Here, we utilized stromal re-engineering to disrupt these barriers and move the stroma toward normalization using a potent antifibrotic agent, halofuginone. In an autochthonous genetically engineered mouse model of PDA, halofuginone disrupted physical barriers to effective drug distribution by decreasing fibroblast activation and reducing key extracellular matrix elements that drive stromal resistance. Concomitantly, halofuginone treatment altered the immune landscape in PDA, with greater immune infiltrate into regions of low hylauronan, which resulted in increased number and distribution of both classically activated inflammatory macrophages and cytotoxic T cells. In concert with a direct effect on carcinoma cells, this led to widespread intratumoral necrosis and reduced tumor volume. These data point to the multifunctional and critical role of the stroma in tumor protection and survival and demonstrate how compromising tumor integrity to move toward a more normal physiologic state through stroma-targeting therapy will likely be an instrumental component in treating PDA.

Author Info: (1) Biomedical Engineering, University of Minnesota. (2) Department of Biomedical Engineering, University of Minnesota. (3) Biomedical Engineering, University of Minnesota. (4) Department of Biomedical Engineering

Author Info: (1) Biomedical Engineering, University of Minnesota. (2) Department of Biomedical Engineering, University of Minnesota. (3) Biomedical Engineering, University of Minnesota. (4) Department of Biomedical Engineering, University of Minnesota pprovenz@umn.edu.

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M2 macrophage-derived exosomes promote cell migration and invasion in colon cancer

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Clinical and experimental evidence has shown that tumor-associated macrophages promote cancer initiation and progression. However, the macrophage-derived molecular determinants that regulate colorectal cancer (CRC) metastasis have not been fully characterized. Here we demonstrate that M2 macrophage-regulated CRC cells migration and invasion is dependent upon M2 macrophage-derived exosomes (MDE). MDE displayed a high expression level of miR-21-5p and miR-155-5p, and MDE-mediated CRC cells migration and invasion depended on these two miRNAs. Mechanistically, miR-21-5p and miR-155-5p were transferred to CRC cells by MDE and bound to the BRG1 coding sequence, downregulating expression of BRG1, which has been identified as a key factor promoting CRC metastasis, yet is downregulated in metastatic CRC cells. Collectively, these findings show that M2 macrophages induce CRC cells migration and invasion and provide significant plasticity of BRG1 expression in response to tumor microenvironments during malignant progression. This dynamic and reciprocal cross-talk between CRC cells and M2 macrophages provides a new opportunity for the treatment of metastatic CRC.

Author Info: (1) Cancer research institute, Tongji Hospital, Tongji Medical College. (2) Department of Oncology, Tongji Hospital, Tongji Medical College. (3) Cancer research institute, Tongji Hospital, Tongji

Author Info: (1) Cancer research institute, Tongji Hospital, Tongji Medical College. (2) Department of Oncology, Tongji Hospital, Tongji Medical College. (3) Cancer research institute, Tongji Hospital, Tongji Medical College. (4) Cancer research institute, Tongji Hospital, Tongji Medical College. (5) Cancer Research Institute, Huazhong University of Science and Technology. (6) Cancer research institute, Tongji Hospital, Tongji Medical College. (7) Cancer Research Institute, Huazhong University of Science and Technology. (8) Cancer research institute, Tongji Hospital, Tongji Medical College. (9) Cancer Research Institute, Huazhong University of Science and Technology. (10) Huazhong University of Science and Technology, Tongji Hospital, Tongji Medical College. (11) Department of Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. (12) Cancer Research Institute, Tongji Hospital, Tongji Medical College. (13) Cancer research institute, Tongji Hospital, Tongji Medical College ghwang@tjh.tjmu.edu.cn.

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Combining DNA vaccine and AIM2 in H1 nanoparticles exert anti-renal carcinoma effects via enhancing tumor-specific multi-functional CD8+ T cell responses

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Renal carcinoma presents a rapid progression in patients with high metastasis with no effective therapeutic strategy. In this study, we designed a folate grafted PEI600-CyD (H1) nanoparticle-mediated DNA vaccine containing an adjuvant of absent in melanoma 2 (AIM2) and a tumor-specific antigen of carbonic anhydrase IX (CAIX) for renal carcinoma therapy. Mice bearing subcutaneous human CAIX (hCAIX)-Renca tumor were intramuscularly immunized with H1-pAIM2/pCAIX, H1-pCAIX, H1-pAIM2, or Mock vaccine, respectively. The tumor growth of hCAIX-Renca was significantly inhibited in H1-pAIM2/pCAIX vaccine group compared with control group. The vaccine activated CAIX-specific CD8+ T cell proliferation and cytotoxic T lymphocyte (CTL) responses, and enhanced the induction of multi-functional CD8+ T cells (expressing TNF-alpha, IL-2, and IFN-gamma). CD8+ T cell depletion resulted in the loss of anti-tumor activity of H1-pAIM2/pCAIX vaccine, suggesting that the efficacy of the vaccine was dependent on CD8+ T cell responses. Lung metastasis of renal carcinoma was also suppressed by H1-pAIM2/pCAIX vaccine treatment accompanied with the increased percentages of CAIX-specific multi-functional CD8+ T cells in the spleen, tumor, and bronchoalveolar lavage as compared with H1-pCAIX vaccine. Similarly, the vaccine enhanced CAIX-specific CD8+ T cell proliferation and CTL responses. Therefore, these results indicated that H1-pAIM2/pCAIX vaccine exhibits the therapeutic efficacy of anti-renal carcinoma by enhancing tumor-specific multi-functional CD8+ T cell responses. This vaccine strategy could be a potential and promising approach for the therapy of primary solid or metastasis tumors.

Author Info: (1) Cancer Institute, Xuzhou Medical University. (2) Department of Orthopedics, Affiliated Hospital of Xuzhou Medical University. (3) Cancer Institute, Xuzhou Medical University. (4) Cancer Institute

Author Info: (1) Cancer Institute, Xuzhou Medical University. (2) Department of Orthopedics, Affiliated Hospital of Xuzhou Medical University. (3) Cancer Institute, Xuzhou Medical University. (4) Cancer Institute, Xuzhou Medical University. (5) Cancer Institute, Xuzhou Medical University. (6) Cancer Institute, Xuzhou Medical University. (7) Cancer Institute, Xuzhou Medical University. (8) Biology college, Hunan University. (9) Cancer Institute, Xuzhou Medical University. (10) Cancer Institute, Xuzhou Medical University. (11) Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute, Xuzhou Medical University jnzheng@xzhmu.edu.cn.

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Collapse of the Plasmacytoid Dendritic Cells compartment in advanced cutaneous melanomas by components of the tumor cell secretome

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Melanoma is an immunogenic neoplasm infiltrated by T cells, although these adaptive T cells usually fail to eradicate the tumor. Plasmacytoid dendritic cells (PDCs) are potent regulators of the adaptive immune response and can eliminate melanoma cells via TLR-mediated effector functions. The PDC compartment is maintained by progressively restricted bone marrow progenitors. Terminally differentiated PDCs exit the bone marrow into the circulation, then home to lymph nodes and inflamed peripheral tissues. Infiltration by PDCs is documented in various cancers. However, their role within the melanoma immune contexture is not completely known. We found that in loco-regional primary cutaneous melanoma (PCM), PDC infiltration was heterogeneous, occurred early, and was recurrently localized at the invasive margin, the site where PDCs interact with CD8+ T cells. A reduced PDC density was coupled with an increased Breslow thickness and somatic mutations at the NRAS p.Q61 codon. Compared to what was seen in PCM, high numbers of PDCs were found in regional lymph nodes, as also identified by in silico analysis. In contrast, in metastatic melanoma (MM) patients, PDCs were mostly absent in the tumor tissues and were significantly reduced in the circulation, particularly in the advanced M1c group. Exposure of circulating PDCs to melanoma cell supernatant (SN-mel) depleted of extracellular vesicles resulted in significant PDC death. SN-mel exposure also resulted in a defect of PDC differentiation from CD34+ progenitors. These findings indicate that soluble components released by melanoma cells support the collapse of the PDC compartment, with clinical implications for refining TLR-agonist based trials.

Author Info: (1) Department of Molecular and Translational Medicine, University of Brescia. (2) Department of Molecular and Translational Medicine, University of Brescia. (3) ASST SPEDALI CIVILI DI

Author Info: (1) Department of Molecular and Translational Medicine, University of Brescia. (2) Department of Molecular and Translational Medicine, University of Brescia. (3) ASST SPEDALI CIVILI DI BRESCIA. (4) Department of Molecular and Translational Medicine, University of Brescia. (5) Medical Oncology, ASST SPEDALI CIVILI DI BRESCIA. (6) DMMT, University of Brescia. (7) Department of Molecular and Translational Medicine, University of Brescia. (8) Department of Molecular and Translational Medicine, Unit of Biostatistics, University of Brescia. (9) Pathology Division, Esine Hospital, ASL Vallecamonica Sebino. (10) Pathology, Azienda USL della Romagna. (11) Department of Molecular and Translational Medicine, University of Brescia. (12) Division of Medical Oncology and Immunotherapy, Department of Oncology, University Hospital of Siena, Istituto Toscano Tumori. (13) Division of Medical Oncology and Immunotherapy, Department of Oncology, University Hospital of Siena, Istituto Toscano Tumori. (14) ASST SPEDALI CIVILI DI BRESCIA. (15) Dep.Diagnostic and Public Health, University and Hospital Trust of Verona. (16) Transfusion Medicine, ASST SPEDALI CIVILI DI BRESCIA. (17) ASST SPEDALI CIVILI DI BRESCIA. (18) Applied Research on Cancer Centre (ARC-Net) and Department of Diagnostics and Public Health-Section of Pathology, University and Hospital Trust of Verona. (19) Department of Molecular and Translational Medicine, University of Brescia. (20) ASST SPEDALI CIVILI DI BRESCIA. (21) Pathology Unit, Department of Molecular and Translational Medicine, University of Brescia. (22) Department of Molecular and Translational Medicine, University of Brescia william.vermi@unibs.it.

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Calnexin impairs the antitumor immunity of CD4+ and CD8+ T cells

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Elucidation of the mechanisms of T cell-mediated antitumor responses will provide information for the rational design and development of cancer immunotherapies. Here, we found that calnexin, an endoplasmic reticulum (ER) chaperone protein, is significantly upregulated in oral squamous cell carcinoma (OSCC). Upregulation of its membranous expression on OSCC cells is associated with inhibited T-cell infiltration in tumor tissues and correlates with poor survival of OSCC patients. We found that calnexin inhibits the proliferation of CD4+ and CD8+ T cells isolated from the whole blood of healthy donors and OSCC patients and inhibits the secretion of IFNgamma, TNFalpha, and IL2 from these cells. Furthermore, in a melanoma model, knockdown of calnexin enhanced the infiltration and effector functions of T cells in the tumor microenvironment and conferred better control of tumor growth, whereas treatment with a recombinant calnexin protein impaired the infiltration and effector functions of T cells and promoted tumor growth. We also found that calnexin enhanced the expression of PD-1 on CD4+ and CD8+ T cells by restraining the DNA methylation status of a CpG island in the PD-1 promoter. Thus, this work uncovers a mechanism by which T-cell antitumor responses are regulated by calnexin in tumor cells and suggests that calnexin might serve as a potential target for the improvement of antitumor immunotherapy.

Author Info: (1) Hospital of Stomatology,Sun yat-sen University. (2) Guanghua School of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Stomatological Hospital, Sun Yat-Sen University. (3) Hospital of

Author Info: (1) Hospital of Stomatology,Sun yat-sen University. (2) Guanghua School of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Stomatological Hospital, Sun Yat-Sen University. (3) Hospital of Stomatology,Sun yat-sen University. (4) Hospital of Stomatology,Sun yat-sen University. (5) Hospital of Stomatology,Sun yat-sen University. (6) Guanghua School of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Stomatological Hospital, Sun Yat-Sen University. (7) Hospital of Stomatology,Sun yat-sen University. (8) Hospital of Stomatology,Sun yat-sen University. (9) Hospital of Stomatology,Sun yat-sen University. (10) Hospital of Stomatology,Sun yat-sen University. (11) Hospital of Stomatology,Sun yat-sen University. (12) Zhongshan School of Medicine, Sun Yat-sen University. (13) Guanghua School of Stomatology, Sun Yat-sen University. (14) College of Life Sciences, Sun Yat-sen University. (15) Zhongshan School of Medicine, Sun Yat-sen University. (16) Immunobiology, Yale University School of Medicine. (17) Guanghua School of Stomatology, Sun Yat-sen University. (18) Guangdong Provincial Key Laboratory of Stomatology, Stomatological Hospital, Sun Yat-Sen University, Guanghua School of Stomatology, Stomatological Hospital, Guangdong Provincial Key Laboratory of Stomatology, SunYat-sen University wangzh75@mail.sysu.edu.cn.

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Clinical immunotherapeutic approaches for the treatment of head and neck cancer

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Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy worldwide, accounting for more than 550,000 cases and 380,000 deaths annually. The primary risk factors associated with HNSCC are tobacco use and alcohol consumption; nevertheless genetic predisposition and oncogenic viruses also play important roles in the development of these malignancies. The current treatments for HNSCC patients include surgery, chemotherapy, radiotherapy, and cetuximab, and combinations of these. However, these treatments are associated with significant toxicity, and many patients are either refractory to the treatment or relapse after a short period. Despite improvements in the treatment of patients with HNSCC, the clinical outcomes of those who have been treated with standard therapies have remained unchanged for over three decades and the 5-year overall survival rate in these patients remains around 40-50%. Therefore, more specific and less toxic therapies are needed in order to improve patient outcomes. The tumour microenvironment of HNSCC is immunosuppressive; therefore immunotherapy strategies that can overcome the immunosuppressive environment and produce long-term tumour immunosurveillance will have a significant therapeutic impact in these patients. This review focuses on the current immunological treatment options under investigation or available for clinical use in patients with HNSCC.

Author Info: (1) Department of Molecular Oncology, King's College London, London, UK. (2) Department of Molecular Oncology, King's College London, London, UK. Electronic address: mahvash.tavassoli@kcl.ac.uk.

Author Info: (1) Department of Molecular Oncology, King's College London, London, UK. (2) Department of Molecular Oncology, King's College London, London, UK. Electronic address: mahvash.tavassoli@kcl.ac.uk.

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Epigenetic regulators of programmed death-ligand 1 expression in human cancers

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The programmed cell death protein 1-programmed death-ligand 1 (PD-L1) axis has been successfully targeted in clinics and the use of immune check-point inhibitors have shown durable antitumor response in untreated or heavily treated advanced stage cancer. PD-L1 upregulation has been found to correlate with poor prognosis in multiple cancer types and expression of PD-L1 in intratumoral compartment has been suggested to influence immune response and act as a key determinant of checkpoint immunotherapy efficacy. Hence it becomes critical to understand the regulation of PD-L1 expression in cancer. Role of oncogenic signaling pathways and transcription factors such as PI3K-AKT, MEK-ERK, JAK-STAT, MYC, HIF-1alpha, AP-1 and NF-kappaB is well established in inducing PD-L1 expression. Even the structural variations resulting in the truncation of the 3' untranslated region (UTR) of PD-L1 has been shown to upregulate PD-L1 expression in multiple cancer types. Since microRNAs carry out post-transcriptional gene silencing by binding to the 3' UTR of its target messenger RNA, truncation of PD-L1 3' UTR can result in alleviation of PD-L1 suppression mediated by microRNA, leading to its overexpression. Other epigenetic modifications, such as promoter DNA methylation and histone modifications can also play crucial role in regulating PD-L1 expression. Here, we review recent findings and evidence on epigenetic mechanisms that regulate PD-L1 expression and the biological and clinical implications of such regulation in cancer.

Author Info: (1) Dept. of Medical Oncology, Dr. B R Ambedkar Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India

Author Info: (1) Dept. of Medical Oncology, Dr. B R Ambedkar Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India. Electronic address: sksingla@gmail.com. (2) Dept. of Medical Oncology, Dr. B R Ambedkar Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.

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Time to abandon single-site irradiation for inducing abscopal effects

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Considerable interest is being directed toward combining immune-checkpoint inhibition (ICI) with radiotherapy to improve response rates to ICI, which have been disappointingly low at around 15-30% among patients with advanced-stage cancers other than melanoma. Since a case report published in 2012, in which authors described the resolution of metastatic disease after irradiation of a single lesion in a patient who had been receiving ICI, hundreds of clinical trials have been launched with the aim of testing the safety and/or efficacy of radiotherapy in combination with immunotherapy, nearly all of which use this single-site irradiation, or 'abscopal', approach. However, emerging preclinical and clinical evidence suggests that this approach likely produces suboptimal results. In this Perspective, we describe this evidence and provide a biological rationale supporting the abandonment of the single-site abscopal approach. We instead advocate exploring comprehensive irradiation of multiple/all lesions in order to enhance the likelihood of obtaining meaningful clinical outcomes - if such a clinical synergy between radiation and ICI does exist - before the failure of the current, single-site approach leads to the potential premature and inappropriate abandonment of radiotherapy in combination with ICI altogether.

Author Info: (1) Department of Radiation Oncology, Unit 97, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (2) Department of Radiation Oncology, Unit 97

Author Info: (1) Department of Radiation Oncology, Unit 97, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (2) Department of Radiation Oncology, Unit 97, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. jychang@mdanderson.org.

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