Construction and self-activation detection of yeast two-hybrid bait plasmid of human programmed cell death ligand 1 immunoglobulin variable region domain gene / 中国生物制品学杂志

@#Objective To construct a yeast two-hybrid recombinant bait plasmid of human programmed cell death ligand 1(PD-L1)immunoglobulin variable region(IgV)domain gene,detect its expression in yeast and detect the cytotoxicity and self-activation of PD-L1 IgV protein as well as the interaction between PD-L1 IgV and human thioredoxin(hTrx).Methods Human PD-L1 was analyzed by bioinformatics method,and primers were designed to amplify PD-L1 IgV domain based on the coding region of PD-L1 gene registered in NCBI GenBank database. PCR amplification was carried out with pENTERPD-L1 plasmid as template,and then cloned into yeast two-hybrid bait vector pGBKT7. The recombinant bait plasmid and pGBKT7 empty vector were transformed into Y2HGold yeast cells respectively,and the PD-L1 IgV gene and its expression were detected by PCR and Western blot;Meanwhile,the protein toxicity and self-activation of PD-L1 IgV were detected,and the interaction between PD-L1 IgV and hTrx was detected by drip plate method.Results The bioinformatics analysis results of PD-L1 were consistent with related reports. The recombinant bait plasmid pGBKT7-PD-L1 IgV was correctly constructed,and Y2HGold positive clone was obtained,in which PD-L1 IgV was stably expressed. The empty vector pGBKT7 and recombinant bait plasmid pGBKT7-PD-L1 IgV grew well on SD/-Trp and SD/-Trp/X-α-Gal plates with the same colony size and number and white colony,but they did not grow on SD/-Trp/X-α-Gal/AbA plates,which indicated that PD-L1 IgV protein had no toxicity and no self-activation effect on yeast. The results of drip plates test showed that all experimental groups grew well on SD/-Trp/-Leu plate,while only positive control group grew on SD/-Trp/-Leu/X-α-Gal/AbA plate and showed blue color,which indicated that bait protein PD-L1 IgV and hTrx did not self-activate,and there was no interaction between them.Conclusion Recombinant human PD-L1 IgV bait plasmid was successfully constructed. PD-L1 IgV protein showed no toxicity and self-activation effect on yeast cells,and there was no interaction between PD-L1 IgV and hTrx. Subsequently,hTrx can be used to construct a peptide aptamer library,from which peptide aptamers that specifically bind to PD-L1 IgV can be screened.

New strategies for targeting PD-1/PD-L1:degraders,bifunctional molecules and covalent inhibitors / 中国药科大学学报

Programmed cell death protein-1(PD-1)/programmed cell death ligand-1(PD-L1)has been considered to be one of the most promising targets for tumor immunotherapy.At present,both monoclonal antibody drugs and small molecule inhibitors targeting PD-1/PD-L1 are facing bottlenecks.Numerous researchers have tried to explore different strategies to block the PD-L1/PD-L1 pathway,hoping to improve the effects of tumor immunotherapy.This review focuses on the degraders,bifunctional molecules and covalent inhibitors that target PD-L1,aiming to provide inspiring insights for the development of anti-PD-1/PD-L1 drugs.

Advances in urinary tract and intestinal microbiome related to bladder cancer / 现代泌尿外科杂志

Bladder cancer is one of the most common urinary tumors, and human urinary tract has been proved to be non-sterile. The significance of urinary tract flora in pathophysiology and treatment of urothelial carcinoma remains to be studied. Meanwhile, intestinal flora has also become an important clinical factor, which has been proved to play a variety of roles in body metabolism, local and systemic inflammation and immunity. Microorganisms can affect the clinical course of cancer, efficacy of chemotherapy and immunotherapy drugs, bioavailability and side effects. This review will explore the relationship between bladder cancer and urinary tract and intestinal microbiome, and explore reliable disease predictors, prognostic indicators and therapeutic targets through a better understanding of the interaction between the microbiome and tumor cells.

Progress on application of PD⁃1／PD⁃L1 monoclonal antibodies in oncotherapy / 中国生物制品学杂志

@#Abstract：Programmed death receptor⁃ 1（PD⁃1）belongs to the family of immunoglobulin B7⁃CD28，which plays an important role in regulating immune response in human body. Since the first PD⁃1／PD⁃ligand 1（PD⁃L1）monoclonal antibody was approved for marketing in China in 2018，the value of PD⁃1／PD⁃L1 immunotherapy in oncotherapy has attracted wide attention. Based on the introduction of the action mechanism of PD⁃1／PD⁃L1 mAbs，this paper reviews the application progress of 8 on ⁃ market PD ⁃ 1／PD ⁃ L1 mAbs in China in oncotherapy from the perspectives of approved indications，clinical trials，usage and dosage，and adverse reactions，in order to provide reference for the rational appli⁃ cation of PD⁃1／PD⁃L1 monoclonal antibodies in clinic.

Advances in tumor immunotherapy / 国际生物医学工程杂志

Tumor immunotherapy has become the main treatment method for cancer after surgery, radiotherapy and chemotherapy. With the in-depth study of tumor immunology, cell biology and molecular technology, the tumor microenvironment was found to be immunosuppressive, and tumor development and metastasis are closely related to it. Tumor immunotherapy uses the body’s own immune system to kill tumor cells. At present, there are mainly mainstream tumor immunotherapy methods, namely monoclonal antibody therapy, immune checkpoint inhibitor therapy, immunocell therapy, and tumor vaccines. In this paper, the advantages and limitations of these four immunotherapies were mainly discussed, and the current status of their marketing and clinical research was also reviewed.

Research progress on dual blockade of TIGIT and PD-1/PD-L1 pathways in tumor im-munotherapy / 中国肿瘤临床

Programmed death-1 and programmed death-ligand 1(PD-1/PD-L1)are regulatory immune checkpoint molecules that inhibit T cell activation and,therefore,play an important role in tumor immunotherapy.In recent years,increasing numbers of targeted therapeutic agents have been developed,but single immune checkpoint blockers cannot completely inhibit tumor occurrence,and tumor escape sporadically occurs.Consequently,combination therapy of targeted drugs is considered a useful method to inhibit tumorigenesis and tumor development.T cell immunoglobulin and immunoreceptor tyrosine-based inhibition motif(ITIM)domain(TIGIT)is an inhibitory type 1 poliovirus receptor that has recently been a hotspot of targeted drug therapy research.It has been shown that the combination therapy of TIGIT plus PD-1/PD-L1 can reduce tumor escape and inhibit tumorigenesis more effectively.Therefore,this review summarizes and discusses the progress on the dual blockade of TIGIT and PD-1/PD-L1 pathways in tumor immunotherapy to provide a theoretical basis for tumor im-munotherapy.

Tumor-associated macrophages in the pancreatic cancer microenvironment: potential therapeutic targets / 中华肝胆外科杂志

Pancreatic cancer is one of the deadliest gastrointestinal malignancies in the world. Although the standard of treatment has been improved, it is not enough to effectively improve the prognosis of patients. As a major component of tumor microenvironment, tumor-associated macrophage (TAM) plays an important role in the field of tumor development, invasion and metastasis, angiogenesis, chemotherapy resistance and immune escape, and are closely related to poor prognosis of patients. Therefore, in-depth exploration of the complex mechanism of TAM in pancreatic cancer immunotherapy may provide new insights into the regulation of pancreatic cancer microenvironment. This article comprehensively introduces the role and mechanism of TAM in the progression of pancreatic cancer, summarizes the pancreatic cancer treatment strategies for TAM and proposes further optimization schemes, hoping to provide a new direction and reference for TAM-based pancreatic cancer immunotherapy.

Progress in the application of pathogen vaccine in tumor immunotherapy / 肿瘤

Immunosuppressive tumor microenvironment is an important obstacle to tumor immunotherapy.Therefore,improving tumor microenvironment to enhance immune response is the key to improve the efficacy of immunotherapy.Based on the basic principle of immunology that the body has a strong response to"foreign"antigens and a weak response to preexisting antigens,pathogen infection can be combined with innate immune-related receptors to enhance the anti-tumor immune response.Pathogen vaccines can induce a"hot"tumor microenvironment with stronger antigen presentation and T lymphocyte activation,showing higher response to immunotherapy and having better security.In this review,we summarized the related studies on pathogen vaccine enhancing tumor immune response,including mixed bacterial vaccine,BCG vaccine,OK-432,synthetic vaccines based on tetanus toxin,modified vaccinia virus,influenza vaccine,Epstein-Barr virus and COVID-19 vaccine.The feasibility,application prospect and challenge of pathogen vaccine in tumor immunotherapy were discussed.

Expressions of long non-coding RNA LINC00673 and ISG15 protein in pancreatic cancer and their clinical significances / 肿瘤研究与临床

Objective:To explore the expressions of long non-coding RNA LINC00673 and ISG15 protein in pancreatic cancer and their clinical significances.Methods:The clinical data of 57 patients diagnosed as pancreatic ductal carcinoma (PDAC) at the Affiliated Cancer Hospital of Xiangya Medical College of Central South University from January 2014 to December 2018 were retrospectively analyzed. The relative expressions of LINC00673 in pancreatic cancer tissues and paracancerous normal tissues (within 3 cm from the edge of cancer tissues) were examined by using quantificational reverse transcription-polymerase chain reaction (qRT-PCR). The ISG15 protein expressions in pancreatic cancer tissues and paracancerous normal tissues were examined by using immunohistochemistry. The difference in LINC00673 expression between ISG15 protein positive and negative patients was compared. The correlation between LINC00673 and ISG15 protein expressions in pancreatic cancer was analyzed by Spearman rank correlation analysis. Moreover, the correlations of LINC00673 and ISG15 protein expressions with clinical stage and pathological classification of pancreatic cancer patients were analyzed.Results:The positive expression of ISG15 protein in pancreatic cancer tissues was 40.4% (23/57), which was higher than that in paracancerous normal tissues [15.8% (9/57)] ( χ2 = 7.90, P = 0.004), and the relative expression of LINC00673 in pancreatic cancer tissues was 0.99±0.36, which was lower than that in paracancerous normal tissues (1.26±0.41) ( t = 4.80, P < 0.001). For 23 (40.4%) ISG15-positive patients and 34 (59.7%) ISG15-negative patients, the relative expression of LINC00673 was 0.77±0.46 and 0.45±0.27 ( P < 0.001). Spearman analysis showed that there was a correlation between LINC00673 and ISG15 protein expressions ( ρ = -0.429, P = 0.001). The relative expression of LINC00673 decreased in patients with low differentiated or undifferentiated tumor, vascular invasion and lymph node metastasis (all P < 0.05), but there was no correlation between LINC00673 expression and patients' age, tumor site, preoperative CA199 level, and TNM stage (all P > 0.05); ISG15 protein expression increased in patients with low differentiated or undifferentiated tumor, TNM stage Ⅲ-Ⅳ, vascular invasion and lymph node metastasis (all P < 0.05), but there was no correlation between ISG15 protein expression and patients' gender, age, tumor site, and preoperative CA199 level (all P > 0.05). Conclusions:The expression of LINC00673 in pancreatic cancer is related to vascular invasion, tumor differentiation degree and lymph node metastasis, and the expression of ISG15 in pancreatic cancer is related to vascular invasion, tumor differentiation degree, lymph node metastasis and TNM stage. The combined detection of LINC00673 and ISG15 protein could be a valuable prognostic indicator for pancreatic cancer. The therapies targeting LINC00673 and ISG15 protein signaling pathways are expected to be a potential option for immunotherapy of pancreatic cancer.

Recent advance in role of cyclic GMP-AMP synthase-stimulator of interferon gene signal pathway in brain tumors / 中华神经医学杂志

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon gene (STING) signal pathway is a classical pathway of STING activation, and in recent years, its role in stimulating innate immunity has gradually attracted wide attention. Besides, cGAS can recognize and combine endogenous or exogenous DNA, then catalyze ATP and GTP to synthesize cyclic guanosine monophosphate-adenosine monophosphate (cGAMP), subsequently activate STING signal to promote type I interferon and inflammatory factors, finally induce natural and adaptive immunity. Existing studies have indicated that cGAS-STING signal pathway plays an important role in infections, inflammations and tumors, especially in high-grade gliomas with poor clinical treatment efficacy. Here, we briefly summarize the cGAS-STING signal pathway and its mechanism in brain tumors to provide new ideas for exploring therapeutic targets and drugs for brain tumors.

Prospect of Tumor Treatment in the Post-PD-1/PD-L1 Inhibitor Era / 肿瘤防治研究

Immune checkpoint inhibitors restart and maintain cancer-immunity circulation to normalize the anti-tumor immunity. Currently, anti-PD-1/PD-L1 antibodies, as new milestone in immunotherapy, have significantly improved the prognosis of patients with various malignant tumors. However, anti-PD-1/PD-L1 antibody alone exhibited a low response rate, and the combination of anti-PD-1/PD-L1 antibody with traditional therapies such as surgery, chemotherapy, radiotherapy and targeted therapy have shown great potential. As new immune checkpoint inhibitors or in combination therapy are on the way, tumor immunotherapy is entering the era of post-anti-PD-1/PD-L1 antibody. The methodology of combination therapy and biomarker screening remain the focus. This paper reviews the current status of immune checkpoint inhibitor therapy and makes a perspective for the future of post-anti-PD-1/PD-L1 antibody era.

Research progress on stimulator of interferon genes (STING) agonists / 中国药科大学学报

@#Stimulator of interferon genes (STING) is an important factor in the auto-immune response of our bodies.Considering the mechanism of activating CD8+ T cells after the activation of STING protein, the combination of STING agonists and immune checkpoint inhibitors for the treatment of tumor immunotherapy has good clinical application prospect.In this paper, the research progress of molecular types, mechanism of action and structural modifications of STING agonists were reviewed.The developing tendency were outlined to provide some references for further investigation.

Future of Immune Checkpoint Therapy for Cancer / 肿瘤防治研究

In 2011, the FDA approved ipilimumab, the first immune checkpoint inhibitor(ICI), targeting CTLA-4, opening the field of immune checkpoint therapy (ICT). ICIs can induce durable clinical responses and improve survival in selected population. However, significant challenges still remain, including mechanisms of resistance, patient selection, management of serious immune-related adverse events, and rational therapeutic combinations. This review surveys the current understanding of response and resistance to ICIs and proposes a path forward to improving efficacy and minimizing toxicities.

Analysis of interactions of immune checkpoint inhibitors with antibiotics in cancer therapy / 医学前沿

The discovery of immune checkpoint inhibitors, such as PD-1/PD-L1 and CTLA-4, has played an important role in the development of cancer immunotherapy. However, immune-related adverse events often occur because of the enhanced immune response enabled by these agents. Antibiotics are widely applied in clinical treatment, and they are inevitably used in combination with immune checkpoint inhibitors. Clinical practice has revealed that antibiotics can weaken the therapeutic response to immune checkpoint inhibitors. Studies have shown that the gut microbiota is essential for the interaction between immune checkpoint inhibitors and antibiotics, although the exact mechanisms remain unclear. This review focuses on the interactions between immune checkpoint inhibitors and antibiotics, with an in-depth discussion about the mechanisms and therapeutic potential of modulating gut microbiota, as well as other new combination strategies.

Research progress of mitogen-activated protein kinase interacting kinases inhibitors in tumor immunotherapy / 中国药科大学学报

@#With the rise of tumor immunotherapy, small molecule modulators targeting the immune system have become a research hotspot. As well-developed and mature targets, immunity protein kinases have attracted more and more attention. Mitogen-activated protein kinase (MAPK)-interacting kinases (MNKs) are located at the meeting-point of ERK and p38 MAPK signaling pathways, which can phosphorylate eukaryotic translation initiation factor 4E (eIF4E) at the conserved serine 209 exclusively and modulate the translation of mRNA. Herein we review the structural characteristics, mechanism, signaling transduction pathways and close relationship with tumors of MNKs.Meanwhile, the development process and clinical research progress of the MNKs inhibitors reported by different research institutions are introduced in detail.

Research Progress of Circular RNAs in Tumor Immunotherapy / 中国肺癌杂志

Tumor immunotherapy is a new therapy which developed in recent years, it has greatly changed the therapeutic schedules and brought new options for patients. However, not all patients can have obvious therapeutic effects after using immunotherapy. So selecting more suitable patients and raising immunotherapy effect are worthy to discuss. With the research of circular RNAs (circRNAs), circRNAs have been found that they not only play a significant role in the field of tumor markers, tumor progression and prognosis, but also can abnormally express in a variety of tumors and affect tumor immunity. Therefore, the circRNAs expression may not only can be used as a supplementary method for selecting patients, but also can be used to predict the efficacy of tumor immunotherapy. In this article, we summarize current knowledge on circRNAs abnormally expressed in many cancers, especially lung cancer which can affect tumor immunity, and discuss its potential effects in tumor immunotherapy, and we hope to provide more references for the clinical practice of circRNAs.
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Research progress on the role of immune cells in tumor hematogenous metastasis / 中国药理学通报

Tumor blood and lymphatic vessel growth are necessary conditions for tumor growth, progression and metastasis, which are closely related with infiltrating nnate as well as adaptive immune cells. On one hand, immune cells rely on adhesion molecules on vascular endothelial cells to penetrate into tumor tissues and show anti-tumor characteristics. On the other hand, they can regulate tumor angiogenesis and lymphangiogenesis by secreting chemokines and cytokines, playing an important role in the process of tumor blood metastasis. In addition, the role of immune cells in promoting tumor blood metastasis highlights the shortcomings of t anti-angiogenic therapy. Therefore, targeting immune cells and tumor angiogenesis improve the effect of therapeutic. Based on these, this article summarizes the effects of immune cells on blood and lymphatic vessels n tumor metastasis, as well as related anti-vascular and immune therapies, in order o provide deas for subsequent research.

Progress in Metabolic Reprogramming of Tumor-associated Macrophages in Tumor Microenvironment / 中国实验方剂学杂志

Occupying more than half of the tumor volume in a variety of solid tumors, tumor-associated macrophages (TAMs) are an important part of the tumor microenvironment (TME) with high plasticity and heterogeneity. In the early stages of tumor development, TAMs mediate antitumor effect through phagocytosis and their antioxidant functions. However, in order to meet the needs of self-renewal and proliferation, malignant tumor cells continuously adjust their metabolic patterns, leading to the accumulation of metabolites such as lactate, reactive oxygen species, nitric oxide, arachidonic acid and prostaglandin in the TME, which results in the changes in its inflammatory profiles, thereby altering the metabolism and function of TAMs and ultimately promoting the tumor development. Therefore, further understanding of the metabolism and immune responses of TAMs in the TME during tumor progression is warranted and the investigation may lead to identification of novel potential targets for cancer immunotherapy. This review aims to clarify the close relationship between TAMs metabolism and TME immune response, to reveal the mechanism of tumor immunosuppression produced by TAMs metabolism, and to provide new treatment ideas and approaches for tumor immunotherapy.

Present status and thought on tumor immunotherapy and related biomarkers / 中国肿瘤临床

Tumor immunotherapy has changed mode for tumor treatment and has gradually developed into the fourth tumor treatment option after surgery, chemotherapy, and radiotherapy. However, the response rate of immunotherapy is limited. Identification of accurate tumor immunotherapy biomarkers as targets or detection and evaluation indicators would be helpful to improve the response rate. Explorations of more effective delivery of immunotherapy would benefit more patients. Prevention and response to immune-related adverse reactions are also necessary. With the development of research, immunotherapy will bring more hope to cancer patients.

EGFR-CAR-T cells secreting PD-1 antibody significantly suppress the progression of gastric cancer / 中国肿瘤生物治疗杂志

@#[Abstract] Objective:To explore the anti-tumor effect of CAR-T cells secreting PD-1 scFv on gastric cancer. Method: We selected EGFR as the target of CAR-T cells and constructed second-generation EGFR-CAR-T cells (EGFR BB-z) and fourth-generation EGFR-CAR-T cells secreting PD-1 scFv (EGFR BB-z/E30). The anti-tumor activity was examined after in vitro activation and long-term stimulation, and its tumor suppression ability was validated through a mouse gastric cell xenograft model. Results: EGFR was highly expressed in gastric cancer tissues and cells (all P<0.01). EGFR BB-z and EGFR BB-z/E30 cells were successfully obtained by lentivirus infection. In vitro experiments showed that compared with EGFR BB-Z, EGFR BB-Z/E30 had longer long-term proliferation ability and stronger tumor killing activity (all P<0.01). In vivo experiments also validated that EGFR BB-z/E30 had obvious tumor inhibitory function in subcutaneous gastric tumor cell transplanted xenograft model and patient-derived tumor xenograft model (PDX) (all P<0.01). It also significantly increased T cell infiltration in tumor site and decreased the expression level of PD-1 (P<0.01 or P<0.05) on EGFR BB-z/E30 cell surface as well as the high secretion of IFN-γ (P<0.05). Conclusion: EGFR-CAR-T cell EGFR BB-z/E30 secreting PD-1 scFv can significantly inhibit the progression of gastric cancer and provide a potential new strategy for the treatment of gastric cancer.