Combined Autophagy Inhibition and Dendritic Cell Recruitment Induces Antitumor Immunity and Enhances Immune Checkpoint Blockade Sensitivity in Pancreatic Cancer.

The effect of immune checkpoint inhibitors is extremely limited in patients with pancreatic ductal adenocarcinoma (PDAC) due to the suppressive tumor immune microenvironment (TIME). Autophagy, which has been shown to play a role in anti-tumor immunity, has been proposed as a therapeutic target for PDAC. Here, single-cell RNA-sequencing of autophagy-deficient murine PDAC tumors revealed that autophagy inhibition in cancer cells induced dendritic cell (DC) activation. Analysis of human PDAC tumors substantiated a negative correlation between autophagy and DC activation signatures. Mechanistically, autophagy inhibition increased intracellular accumulation of tumor antigens, which could activate DCs. Administration of chloroquine (CQ), an autophagy inhibitor, in combination with Flt3 ligand (Flt3L)-induced DC infiltration inhibited tumor growth and increased tumor-infiltrating T lymphocytes. However, autophagy inhibition in cancer cells also induced CD8+ T cell exhaustion with high expression of immune checkpoint LAG3. A triple therapy comprising CQ, Flt3L, and an anti-LAG3 antibody markedly reduced tumor growth in orthotopic syngeneic PDAC mouse models. Thus, targeting autophagy in cancer cells and activating DCs sensitizes PDAC tumors to immune checkpoint inhibitor therapy, warranting further development of this treatment approach to overcome immunosuppression in pancreatic cancer.

Tumor-associated neutrophils upregulate Nectin2 expression, creating the immunosuppressive microenvironment in pancreatic ductal adenocarcinoma.

BACKGROUND: Tumor-associated neutrophils (TANs) constitute an abundant component among tumor-infiltrating immune cells and have recently emerged as a critical player in pancreatic ductal adenocarcinoma (PDAC) progression. This study aimed to elucidate the pro-tumor mechanisms of TAN and identify a novel target for effective immunotherapy against PDAC. METHODS: Microarray and cytokine array analyses were performed to identify the mechanisms underlying the function of TANs. Human and mouse TANs were obtained from differentiated HL-60 cells and orthotopically transplanted PDAC tumors, respectively. The interactions of TANs with cancer and cytotoxic T-cells were evaluated through in vitro co-culture and in vivo orthotopic or subcutaneous models. Single-cell transcriptomes from patients with PDAC were analyzed to validate the cellular findings. RESULTS: Increased neutrophil infiltration in the tumor microenvironment was associated with poor survival in patients with PDAC. TANs secreted abundant amounts of chemokine ligand 5 (CCL5), subsequently enhancing cancer cell migration and invasion. TANs subpopulations negatively correlated with cytotoxic CD8+ T-cell infiltration in PDAC and promoted T-cell dysfunction. TANs upregulated the membranous expression of Nectin2, which contributed to CD8+ T-cell exhaustion. Blocking Nectin2 improved CD8+ T-cell function and suppressed tumor progression in the mouse model. Single-cell analysis of human PDAC revealed two immunosuppressive TANs phenotypes: Nectin2+ TANs and OLR1+ TANs. Endoplasmic reticulum stress regulated the protumor activities in TANs. CONCLUSIONS: TANs enhance PDAC progression by secreting CCL5 and upregulating Nectin2. Targeting the immune checkpoint Nectin2 could represent a novel strategy to enhance immunotherapy efficacy in PDAC.

Blockade of histamine receptor H1 augments immune checkpoint therapy by enhancing MHC-I expression in pancreatic cancer cells.

BACKGROUND: Although immune checkpoint blockade (ICB) therapy has proven to be extremely effective at managing certain cancers, its efficacy in treating pancreatic ductal adenocarcinoma (PDAC) has been limited. Therefore, enhancing the effect of ICB could improve the prognosis of PDAC. In this study, we focused on the histamine receptor H1 (HRH1) and investigated its impact on ICB therapy for PDAC. METHODS: We assessed HRH1 expression in pancreatic cancer cell (PCC) specimens from PDAC patients through public data analysis and immunohistochemical (IHC) staining. The impact of HRH1 in PCCs was evaluated using HRH1 antagonists and small hairpin RNA (shRNA). Techniques including Western blot, flow cytometry, quantitative reverse transcription polymerase chain reaction (RT-PCR), and microarray analyses were performed to identify the relationships between HRH1 and major histocompatibility complex class I (MHC-I) expression in cancer cells. We combined HRH1 antagonism or knockdown with anti-programmed death receptor 1 (αPD-1) therapy in orthotopic models, employing IHC, immunofluorescence, and hematoxylin and eosin staining for assessment. RESULTS: HRH1 expression in cancer cells was negatively correlated with HLA-ABC expression, CD8+ T cells, and cytotoxic CD8+ T cells. Our findings indicate that HRH1 blockade upregulates MHC-I expression in PCCs via cholesterol biosynthesis signaling. In the orthotopic model, the combined inhibition of HRH1 and αPD-1 blockade enhanced cytotoxic CD8+ T cell penetration and efficacy, overcoming resistance to ICB therapy. CONCLUSIONS: HRH1 plays an immunosuppressive role in cancer cells. Consequently, HRH1 intervention may be a promising method to amplify the responsiveness of PDAC to immunotherapy.

Intratumor Fusobacterium nucleatum promotes the progression of pancreatic cancer via the CXCL1-CXCR2 axis.

Intratumor bacteria modify the tumor immune microenvironment and influence outcomes of various tumors. Periodontal pathogen Fusobacterium nucleatum has been detected in pancreatic cancer tissues and is associated with poor prognosis. However, it remains unclear how F. nucleatum affects pancreatic cancer. Here, we compared clinical features with F. nucleatum colonization in pancreatic cancer tissues. F. nucleatum was detected in 15.5% (13/84) of pancreatic cancer patients. The tumor size was significantly larger in the F. nucleatum-positive group than in the negative group. To clarify the biological effect of intratumor F. nucleatum on pancreatic cancer progression, we performed migration/invasion assays and cytokine array analysis of cancer cells cocultured with F. nucleatum. F. nucleatum promoted CXCL1 secretion from pancreatic cancer cells, leading to cancer progression through autocrine signaling. Intratumor F. nucleatum suppressed tumor-infiltrating CD8+ T cells by recruiting myeloid-derived suppressor cells (MDSCs) to the tumor in an F. nucleatum-injected subcutaneous pancreatic cancer mouse model, resulting in tumor progression. Furthermore, tumor growth accelerated by F. nucleatum was suppressed by MDSC depletion or cytokine inhibitors. Intratumor F. nucleatum promoted pancreatic cancer progression through autocrine and paracrine mechanisms of the CXCL1-CXCR2 axis. Blockade of the CXCL1-CXCR2 axis may be a novel therapeutic approach for patients with intratumor F. nucleatum-positive pancreatic cancer.

Novel insights into the relationship between α-1 anti-trypsin with the pathological development of cardio-metabolic disorders.

According to the previous studies, chronic low-grade systemic inflammatory response has been shown to be significantly associated with the pathological development of cardio-metabolic disorder diseases, including atherosclerosis, type 2 diabetes mellitus, and non-alcoholic fatty liver disease (NAFLD). On the other hand, auto-immunity process could also facilitate the pathogenesis of type 1 diabetes mellitus importantly. Concerning on this notion, the anti-inflammatory therapeutic strategy is demonstrated to embrace an essential function in those cardio-metabolic disorders in clinical practice. The α-1 anti-trypsin, also named Serpin-A1 and as an acute phase endogenous protein, has been verified to have several modulatory effects such as anti-inflammatory response, anti-apoptosis, and immunomodulatory functions. In addition, it is also used for therapeutic strategy of a rare genetic disease caused by the deficiency of α-1 anti-trypsin. Recent emerging evidence has indicated that the serum concentrations of α-1 anti-trypsin levels and its biological activity are significantly changed in those inflammatory and immune related cardio-metabolic disorder diseases. Nevertheless, the underlying mechanism is still not elucidated. In the current review, the basic experiments and clinical trials which provided the evidence revealing the potential therapeutic function of the α-1 anti-trypsin in cardio-metabolic disorder diseases were well-summarized. Furthermore, the results which indicated that the α-1 anti-trypsin presented the possibility as a novel serum biomarker in humans to predict those cardio-metabolic disorder diseases were also elucidated.

A Study on the Relationship between Postural Control and Pain-Related Clinical Outcomes in Patients with Chronic Nonspecific Low Back Pain.

OBJECTIVES: To explore the relationship between postural control and pain-related clinical outcomes in patients with chronic nonspecific low back pain (cNLBP). METHODS: Participants with cNLBP and healthy individuals were recruited. Muscle activities were recorded during internal and external perturbation tasks. Postural control capacity was assessed by muscle onset time and integrals of electromyography (iEMGs) of postural muscles during the phases of anticipatory postural adjustments (APAs) and compensatory postural adjustments (CPAs). Correlation analysis was employed to investigate the relationship between postural control capacity, pain, and disability. RESULTS: Twenty-seven patients with cNLBP and 27 healthy participants were recruited. Gastrocnemius (GA) muscle onset time was earlier in the cNLBP group than in the control group in the internal perturbation task. The onset time of GA and erector spinae (ES) of the cNLBP group was later than that of the controls in the external perturbation task. Disability level moderately correlated with the iEMGs of rectus abdominis (RA), GA, and external oblique (EO) during APAs. Pain score moderately correlated with the iEMGs of RA, EO, and ES during CPAs of perturbation tasks. CONCLUSION: cNLBP participants had altered muscle activation strategy to maintain postural stability in response to perturbation. This study further discovered that pain-related disabilities of cNLBP participants were likely related to the APAs capacity, whereas the pain intensity may relate to the CPAs capacity. Pain and disability may therefore be related to the control process of the posture-related muscles.

The Effect of Virtual Reality Training on Anticipatory Postural Adjustments in Patients with Chronic Nonspecific Low Back Pain: A Preliminary Study.

Objectives: This study is aimed at exploring the effects of virtual reality (VR) training on postural control, measured by anticipatory and compensatory postural adjustments (APAs and CPAs, respectively), in patients with chronic nonspecific low back pain (CNLBP) and the potential neuromuscular mechanism of VR training. Methods: Thirty-four patients were recruited and randomly assigned to the VR group (n = 11), the motor control exercise group (MCE, n = 12) and the control group (CG, n = 11). The VR group received VR training using Kinect Xbox 360 systems and magnetic therapy. Besides magnetic therapy, the participants in the MCE group performed real-time ultrasound-guided abdominal drawing-in maneuver (ADIM) and four-point kneeling exercise. The CG only received magnetic therapy. Surface muscle electromyography (sEMG) was used to record the muscle activities of transverse abdominis (TrA), multifidus (MF), lateral gastrocnemius (LG), and tibialis anterior (TA) during ball-hitting tasks. The muscle activation time and integrals of the electromyography activities (IEMGs) during the APA and CPA stages were calculated and used in the data analysis. The visual analogue scale (VAS) and Oswestry dysfunction index (ODI) scores were also recorded. Results: A significant interaction effect of time × group was observed on the activation time of TrA (p = 0.018) and MF (p = 0.037). The post-intervention activation time of the TrA was earlier in the VR group (p = 0.029). In contrast, the post-intervention activation time of the MF was significantly delayed in the VR group (p = 0.001). The IEMGs of TrA (p = 0.002) and TA (p = 0.007) during CPA1 significantly decreased only in the VR group after the intervention. The VAS scores of three group participants showed significant decreases after intervention (p < 0.001). Conclusions: Patients with CNLBP showed reciprocal muscle activation patterns of the TrA and MF muscles after VR training. VR training may be a potential intervention for enhancing the APAs of the patients with CNLBP.

The Effect of Visual Stimuli on Stability and Complexity of Postural Control.

Visual input could benefit balance control or increase postural sway, and it is far from fully understanding the effect of visual stimuli on postural stability and its underlying mechanism. In this study, the effect of different visual inputs on stability and complexity of postural control was examined by analyzing the mean velocity (MV), SD, and fuzzy approximate entropy (fApEn) of the center of pressure (COP) signal during quiet upright standing. We designed five visual exposure conditions: eyes-closed, eyes-open (EO), and three virtual reality (VR) scenes (VR1-VR3). The VR scenes were a limited field view of an optokinetic drum rotating around yaw (VR1), pitch (VR2), and roll (VR3) axes, respectively. Sixteen healthy subjects were involved in the experiment, and their COP trajectories were assessed from the force plate data. MV, SD, and fApEn of the COP in anterior-posterior (AP), medial-lateral (ML) directions were calculated. Two-way analysis of variance with repeated measures was conducted to test the statistical significance. We found that all the three parameters obtained the lowest values in the EO condition, and highest in the VR3 condition. We also found that the active neuromuscular intervention, indicated by fApEn, in response to changing the visual exposure conditions were more adaptive in AP direction, and the stability, indicated by SD, in ML direction reflected the changes of visual scenes. MV was found to capture both instability and active neuromuscular control dynamics. It seemed that the three parameters provided compensatory information about the postural control in the immersive virtual environment.