PlexinB1 inactivation reprograms immune cells in the tumor microenvironment, inhibiting breast cancer growth and metastatic dissemination.

Semaphorin-Plexin signaling plays a major role in the tumor microenvironment (TME). In particular, Semaphorin 4D (SEMA4D) has been shown to promote tumor growth and metastasis; however, the role of its high-affinity receptor Plexin-B1 (PLXNB1), which is expressed in the TME, is poorly understood. In this study, we directly targeted PLXNB1 in the TME of triple-negative murine breast carcinoma to elucidate its relevance in cancer progression. We found that primary tumor growth, and metastatic dissemination were strongly reduced in PLXNB1-deficient mice, which showed longer survival. PLXNB1-loss in the TME induced a switch in the polarization of tumor-associated macrophages (TAMs) towards a pro-inflammatory M1 phenotype and enhanced the infiltration of CD8+ T lymphocytes both in primary tumors and in distant metastases. Moreover, PLXNB1-deficiency promoted a shift in the Th1/Th2 balance of the T-cell population and an antitumor gene signature, with the up-regulation of Icos, Perforin-1, Stat3 and Ccl5 in tumor infiltrating lymphocytes (TILs). We thus tested the translational relevance of TME re-programming driven by PLXNB1 inactivation for responsiveness to immunotherapy. Indeed, in the absence of PLXNB1, the efficacy of anti-PD-1 blockade was strongly enhanced, efficiently reducing tumor growth and distant metastasis. Consistent with this, pharmacological PLXNB1 blockade by systemic treatment with a specific inhibitor significantly hampered breast cancer growth and enhanced the antitumor activity of the anti-PD1 treatment in a preclinical model. Altogether, these data indicate that PLXNB1 signaling controls the antitumor immune response in the TME and highlight this receptor as a promising immune therapeutic target for metastatic breast cancers.

Complications in endoscopic spine surgery: a systematic review.

PURPOSE: This systematic review aims to investigate the complication rate of endoscopic spine surgeries, stratifying them by technique, district and kind of procedure performed. METHODS: This study was conducted according to the PRISMA statement. The literature search was conducted in MEDLINE, CINAHL, EMBASE, Cochrane Register, OTseeker and ScienceDirect database. Types of studies included were observational studies (cohort studies, case-control studies and case series) and randomised or quasi-randomised clinical with human subjects. No restrictions on publication year were applied. Repeated articles, reviews, expert's comments, congress abstracts, technical notes and articles not in English were excluded. Several data were extracted from the articles. In particular, data of perioperative (≤ 3 months) and late (> 3 months) complications were collected and grouped according to: (1) surgical technique [uniportal full-endoscopic spine surgery (UESS) or unilateral biportal endoscopic spine surgery (UBESS)]; (2) spinal district treated [cervical, thoracic or lumbar] and (3) type of procedure [discectomy/decompression or fusion]. Complication analysis was performed in subgroups with at least 100 patients to have clinically meaningful statistical validity. RESULTS: A total of 117 full-text articles were assessed for eligibility. Of the 117 records included, 95 focused their research on UESS (14 LOE V, 33 LOE IV, 43 LOE III and five LOE II) and 23 on UBESS (three LOE V, eight LOE IV, 10 LOE III and two LOE II). A total of 20,020 patients were extracted to investigate the incidence of different perioperative and late complications, 10,405 for UESS and 9615 for UBESS. CONCLUSION: The present study summarises the complications reported in the literature for spinal endoscopic procedures. On the one hand, the most relevant described were perioperative complications (transient neurological deficit, dural tear and dysesthesia) that are especially meaningful for endoscopic discectomy and decompression. On the other hand, late complications, such as mechanical implant failure, are more common in endoscopic interbody fusion. LEVEL OF EVIDENCE: I.

The influence of peri-operative depressive symptoms on medium-term spine surgery outcome: a prospective study.

PURPOSE: To investigate the role of depressive symptoms on clinical outcomes in patients undergoing spinal surgery up to 2-year follow-up. METHODS: The study used data from an institutional spine surgery registry (January 2016, through March 2022) to identify patients (> 18 years) undergoing spine surgery. Patients with Oswestry Disability Index (ODI) < 20/100 at baseline or undergoing surgery on the cervical spine or for idiopathic spinal deformity and trauma patients were excluded. The patients were divided into two groups based on the pre-operative Mental Component Summary (MCS) score of the SF-36: depression group (MCS ≤ 35) or non-depression group (MCS > 35). The ODI and MCS scores trajectory were wined over the 24-month post-surgery between groups. Additionally, a secondary subgroup analysis was conducted comparing outcomes between those with depressive symptoms (persistent-depression subgroup) and those without depressive symptoms (never-depression subgroup) at 3 months after surgery. RESULTS: A total of 2164 patients who underwent spine surgery were included. The pre-operative depression group reported higher ODI total scores and lower MCS than the pre-operative non-depression group at all time points (P < 0.001). The persistent-depression subgroup reported higher ODI total scores and lower MCS than the never-depression subgroup at all follow-ups (P < 0.001). CONCLUSION: Functional disability and mental health status improve in patients with depression symptoms undergoing spinal surgery. Despite this improvement, they do not reach the values of non-depressed subjects. Over the 2-year follow-up time, patients with depression show a different trajectory of ODI and MCS. Caregivers should be aware of these results to counsel patients with depression symptoms.

Oblique, Unilateral, or Bilateral Rods Configurations for Single-Level Interbody Fusion and Posterior Spinal Fixation: A Finite Element Study.

AIM: To compare three different posterior mono-segmental instrumented models with a Lateral Lumbar Interbody Fusion (LLIF) cage in L4-L5 based on finite element (FE) analysis. MATERIAL AND METHODS: Three different configurations of posterior instrumentation were created: 1. Bilateral posterior screws with 2 rods: Bilateral (B); 2. Left posterior rod and left pedicle screws in L4-L5: Unilateral (U); 3. Oblique posterior rod, left pedicle screw in L4, and right pedicle screw in L5: Oblique (O). The models were compared regarding the range of motion (ROM), stresses in the L4 and L5 pedicle screws, and posterior rods. RESULTS: The Oblique and Unilateral models showed a lower decrease in ROM than the Bilateral model (O vs U vs B; 92% vs 95% vs 96%). In the L4 screw, a higher stress level was identified in the O than in the B model. Still, lower if compared to U. In the L5 screw, the highest stress values were observed with the O model in extension and flexion and the U model in lateral bending and axial rotation. The highest stress values for the rods were observed for the O model in extension, flexion, and axial rotation and the U model in lateral bending. CONCLUSION: The FE analysis showed that the three configurations significantly reduced the ROM. The stress analysis identified a substantially higher value for the rod and pedicle screws in oblique or unilateral configuration systems compared to the standard bilateral one. In particular, the oblique configuration has stress properties similar to the unilateral in lateral bending and axial rotation but is significantly higher in flexion-extension.

The burden of preoperative fear-avoidance beliefs in workers after thoracic and lumbar spine surgery: a 2-year follow-up study.

ABSTRACT: Spinal disorders are the main reasons for sick leave and early retirement among the working population in industrialized countries. When "red flags" are present, spine surgery is the treatment of choice. However, the role of psychosocial factors such as fear-avoidance beliefs in spine surgery outcomes is still debated. The study aims to investigate whether patients presenting high or low levels of fear-avoidance thoughts before the spine surgery reported different surgical results and return-to-work rates over 2 years. From an institutional spine surgery registry, workers surgically treated with a preoperative score in the Oswestry Disability Index (ODI) higher than 20/100 and provided ODI questionnaires, return-to-work status at 3-, 6-, 12-, and 24-month follow-ups were analyzed. A total of 1769 patients were stratified according to the work subscale of the Fear-Avoidance Beliefs Questionnaire (FABQ-W) in high fear (FABQ-W ≥ 34/42) or low fear (FABQ-W < 34/42). Multivariate regression was used to search for preoperative factors, which might interact with FABQ-W. The higher-fear group showed a different recovery pattern, with higher levels of disability according to the ODI (total score, absolute change, frequency of clinically relevant change, and disability categories) and lower return-to-work ratios over the 24-month follow-up. High fear, high disability, greater age, female gender, smoking, and worse physical status at baseline were associated with worse ODI outcomes 2 years after the surgery. In summary, fear-avoidance beliefs significantly influence the speed and the entity of surgical outcomes in the working population. However, the contribution of FABQ-W in predicting long-term disability levels was limited.

Correction: Ponzo et al. Nucleolin Therapeutic Targeting Decreases Pancreatic Cancer Immunosuppression. Cancers 2022, 14, 4265.

In the original publication [...].

Nucleolin Therapeutic Targeting Decreases Pancreatic Cancer Immunosuppression.

Background: The pancreatic ductal adenocarcinoma (PDAC) microenvironment is highly fibrotic and hypoxic, with poor immune cell infiltration. Recently, we showed that nucleolin (NCL) inhibition normalizes tumour vessels and impairs PDAC growth. Methods: Immunocompetent mouse models of PDAC were treated by the pseudopeptide N6L, which selectively inhibits NCL. Tumour-infiltrating immune cells and changes in the tumour microenvironment were analysed. Results: N6L reduced the proportion of regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs) and increased tumour-infiltrated T lymphocytes (TILs) with an activated phenotype. Low-dose anti-VEGFR2 treatment normalized PDAC vessels but did not modulate the immune suppressive microenvironment. RNAseq analysis of N6L-treated PDAC tumours revealed a reduction of cancer-associated fibroblast (CAF) expansion in vivo and in vitro. Notably, N6L treatment decreased IL-6 levels both in tumour tissues and in serum. Treating mPDAC by an antibody blocking IL-6 reduced the proportion of Tregs and MDSCs and increased the amount of TILs, thus mimicking the effects of N6L. Conclusions: These results demonstrate that NCL inhibition blocks the amplification of lymphoid and myeloid immunosuppressive cells and promotes T cell activation in PDAC through a new mechanism of action dependent on the direct inhibition of the tumoral stroma.

Nucleolin Targeting by N6L Inhibits Wnt/ß-Catenin Pathway Activation in Pancreatic Ductal Adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive and resistant cancer with no available effective therapy. We have previously demonstrated that nucleolin targeting by N6L impairs tumor growth and normalizes tumor vessels in PDAC mouse models. Here, we investigated new pathways that are regulated by nucleolin in PDAC. We found that N6L and nucleolin interact with ß-catenin. We found that the Wnt/ß-catenin pathway is activated in PDAC and is necessary for tumor-derived 3D growth. N6L and nucleolin loss of function induced by siRNA inhibited Wnt pathway activation by preventing ß-catenin stabilization in PDAC cells. N6L also inhibited the growth and the activation of the Wnt/ß-catenin pathway in vivo in mice and in 3D cultures derived from MIA PaCa2 tumors. On the other hand, nucleolin overexpression increased ß-catenin stabilization. In conclusion, in this study, we identified ß-catenin as a new nucleolin interactor and suggest that the Wnt/ß-catenin pathway could be a new target of the nucleolin antagonist N6L in PDAC.

Tumor stiffening reversion through collagen crosslinking inhibition improves T cell migration and anti-PD-1 treatment.

Only a fraction of cancer patients benefits from immune checkpoint inhibitors. This may be partly due to the dense extracellular matrix (ECM) that forms a barrier for T cells. Comparing five preclinical mouse tumor models with heterogeneous tumor microenvironments, we aimed to relate the rate of tumor stiffening with the remodeling of ECM architecture and to determine how these features affect intratumoral T cell migration. An ECM-targeted strategy, based on the inhibition of lysyl oxidase, was used. In vivo stiffness measurements were found to be strongly correlated with tumor growth and ECM crosslinking but negatively correlated with T cell migration. Interfering with collagen stabilization reduces ECM content and tumor stiffness leading to improved T cell migration and increased efficacy of anti-PD-1 blockade. This study highlights the rationale of mechanical characterizations in solid tumors to understand resistance to immunotherapy and of combining treatment strategies targeting the ECM with anti-PD-1 therapy.

Pharmacologic Normalization of Pancreatic Cancer-Associated Fibroblast Secretome Impairs Prometastatic Cross-Talk With Macrophages.

BACKGROUND & AIMS: Cancer-associated fibroblasts (CAFs) from pancreatic adenocarcinoma (PDA) present high protein synthesis rates. CAFs express the G-protein-coupled somatostatin receptor sst1. The sst1 agonist SOM230 blocks CAF protumoral features in vitro and in immunocompromised mice. We have explored here the therapeutic potential of SOM230, and underlying mechanisms, in immunocompetent models of murine PDA mimicking the heavy fibrotic and immunosuppressive stroma observed in patient tumors. METHODS: Large-scale mass spectrometry analyses were performed on media conditioned from 9 patient PDA-derived CAF primary cultures. Spontaneous transgenic and experimental (orthotopic co-graft of tumor cells plus CAFs) PDA-bearing mice were longitudinally ultrasound-monitored for tumor and metastatic progression. Histopathology and flow cytometry analyses were performed on primary tumors and metastases. Stromal signatures were functionally validated through bioinformatics using several published, and 1 original, PDA database. RESULTS: Proteomics on the CAF secretome showed that SOM230 controls stromal activities including inflammatory responses. Among the identified secreted proteins, we validated that colony-stimulating factor 1 (CSF-1) (a macrophage growth factor) was reduced by SOM230 in the tumor and plasma of PDA-harboring mice, alongside intratumor stromal normalization (reduced CAF and macrophage activities), and dramatic metastasis reduction. In transgenic mice, these SOM230 benefits alleviate the chemotherapy-induced (gemcitabine) immunosuppressive stroma reshaping. Mechanistically, SOM230 acts in vivo on CAFs through sst1 to disrupt prometastatic CAF production of CSF-1 and cross-talk with macrophages. We found that in patients, stromal CSF-1 was associated with aggressive PDA forms. CONCLUSIONS: We propose SOM230 as an antimetastatic therapy in PDA for its capacity to remodel the fibrotic and immunosuppressive myeloid stroma. This pharmacotherapy should benefit PDA patients treated with chemotherapies.

Weibel-Palade Bodies Orchestrate Pericytes During Angiogenesis.

Objective Weibel-Palade bodies (WPBs) are endothelial cell (EC)-specific organelles formed by vWF (von Willebrand factor) polymerization and that contain the proangiogenic factor Ang-2 (angiopoietin-2). WPB exocytosis has been shown to be implicated for vascular repair and inflammatory responses. Here, we investigate the role of WPBs during angiogenesis and vessel stabilization. Approach and Results WPB density in ECs decreased at the angiogenic front of retinal vascular network during development and neovascularization compared with stable vessels. In vitro, VEGF (vascular endothelial growth factor) induced a VEGFR-2 (vascular endothelial growth factor receptor-2)-dependent exocytosis of WPBs that contain Ang-2 and consequently the secretion of vWF and Ang-2. Blocking VEGF-dependant WPB exocytosis and Ang-2 secretion promoted pericyte migration toward ECs. Pericyte migration was inhibited by adding recombinant Ang-2 or by silencing Ang-1 (angiopoietin-1) or Tie2 (angiopoietin-1 receptor) in pericytes. Consistently, in vivo anti-VEGF treatment induced accumulation of WPBs in retinal vessels because of the inhibition of WPB exocytosis and promoted the increase of pericyte coverage of retinal vessels during angiogenesis. In tumor angiogenesis, depletion of WPBs in vWF knockout tumor-bearing mice promoted an increase of tumor angiogenesis and a decrease of pericyte coverage of tumor vessels. By another approach, normalized tumor vessels had higher WPB density. Conclusions We demonstrate that WPB exocytosis and Ang-2 secretion are regulated during angiogenesis to limit pericyte coverage of remodeling vessels by disrupting Ang-1/Tie2 autocrine signaling in pericytes.