Glioblastoma (GBM) is the most aggressive and devastating primary brain cancer in adults. Most GBMs are diagnosed at an advanced stage with therapy resistance, posing a major obstacle to understanding the tumor microenvironment (TME) at the earliest stages of disease development. A precise characterization of early-stage GBM and its TME could provide critical insights into tumor progression and inform new therapeutic strategies. In a recent issue of Nature, Clements and colleagues demonstrated that white matter (WM) injury, induced by early tumor cells, constitutes a key TME factor driving GBM progression. Using somatic mouse models, patient-derived xenografts, and human tissues, they showed that early glioma cells preferentially infiltrate WM tracts, inducing sterile alpha and TIR motif-containing 1-mediated Wallerian degeneration that propagates into distal WM regions. Remarkably, WM injury induced by axonal transection significantly accelerated GBM progression at distal sites, whereas this effect was abolished by Sarm1 knockout, confirming that axonal injury followed by Wallerian degeneration drives distal tumor progression. Collectively, these findings reveal a previously unrecognized evolutionary process in GBM development and highlight potential targets for therapeutic intervention.

Legends to Supplementary Figures 1-6

Supplemental Table 1: Patient Demographics.

PDF file 106K, Table S1. Molecules whose expression significantly differed between cells treated with KX-01 and control cells in RMUG-S cells Table S2. Molecules whose expression significantly differed between cells treated with KX-01 and control cells in RMUG-L cells

Supplementary Figure 5. MGMT promoter methylation in orthotopically implanted LN- 229, LN-229Res, and D54Res glioblastomas.

Supplementary Figure 6. Effects of treatment on glioblastoma-associated blood vessels.

Purpose: Antibiotic use preceding immune checkpoint inhibitor (ICI) treatment has been associated with a decreased efficacy of ICI in solid tumors. In this study, we evaluated the effect of antibiotic use before ICI therapy on oncological outcomes. Methods: We examined patients with recurrent gynecologic malignancies at two academic institutions. The clinical data, including antibiotic use within 60 days of ICI initiation, type of antibiotics, reasons for antibiotic use, body mass index, tumor site, chemotherapy-free interval, prior history of radiotherapy, disease control rate (DCR), and overall survival (OS), were assessed. Results: Of 215 patients, 22.9% (n = 47) received antibiotics before ICI treatment. The most common cancer was ovarian (52.1%, n = 112), followed by cervical (24.7%, n = 53) and endometrial (16.7%, n = 36). When we divided the cohort based on antibiotic use before ICIs, there were no significant differences in the DCR and baseline characteristics between the two groups. On multivariate analyses, the variables associated with poor OS were previous use of antibiotics for a cumulative duration of >14 days (HR 2.286, 95% CI 1.210-4.318; p = 0.011); Eastern Cooperative Oncology Group 2 or 3 (HR 4.677, 95% CI 2.497-8.762; p < 0.001); and chemotherapy-free interval of <6 months (HR 2.007, 95% CI 1.055-3.819; p = 0.034). Conclusion: Prior use of antibiotics for a cumulative duration of >14 days was associated with reduced survival in recurrent gynecologic malignancies.

PDF file 86K, Figure S7. (A) Western blot results showing PTEN expression in RMUG-S and RMUG-L cells. PTEN expression is null in the RMUG-L cells. (B) MTT assay results showing cytotoxicity of an Akt1/2 inhibitor in RMUG-S and RMUG-L cells exposed to various concentrations of Akt1/2 inhibitor for 72 hours. Cell survival was calculated as the number of cells surviving relative to the number of cells surviving in the control group. (C) Western blot results showing endogenous p-Akt expression levels in RMUG-S and RMUG-L cells after treatment with 15 uM of Akt1/2 inhibitor

Supplementary Figure 2. Combination therapy with macitentan plus TMZ eradicates experimental glioblastomas.

Objectives: Antibiotic use or BMI has been reported to be associated with the effectiveness of ICI in solid tumors. We evaluated associations between antibiotic use either prior to ICI or during ICI, BMI at ICI initiation, and oncological outcomes. Methods: We examined patients with recurrent gynecologic malignancies who underwent PD-1 monotherapy at two academic institutions. Clinical data, including antibiotic use within 30 days of initiation of ICI or during ICI treatment, BMI, tumor site, chemotherapy-free interval, prior history of radiotherapy, the objective response at 12 weeks, PFS, and OS, were assessed. Results: A total of 204 patients were eligible, of whom 22.1% (45/204) and 41.2% (84/204) received antibiotics before and during ICI, respectively. Among the patients, 18.1% (37/204) were underweight. Based on tumor site, ovarian cancer was most common (51.0%, 104/204), followed by the cervix (24.5%, 50/204) and endometrial cancer (17.6%, 36/204). About 45.1% of patients had one or two previous lines of chemotherapy, and 80.4% (164/204) patients had chemotherapy-free intervals of less than six months. In terms of overall survival, endometrial cancer showed the best survivals among gynecologic malignancies (p = 0.034). In addition, low BMI, antibiotic use before or during ICI, and chemotherapy-free interval were associated with poor overall survival. In multivariate analyses, the negative association between low BMI (HR: 2.209; 95% CI: 1.236-3.949; p=0.007) and antibiotic use during ICI (HR: 1.910; 95% CI: 1.106-3.297; p=0.02) remained significant for OS. No significant association between objective response rates at 12 weeks and BMI or antibiotics use was observed. Conclusions: Antibiotics use during ICI, or low BMI were associated with reduced survival from ICI in recurrent gynecologic malignancies, which is in line with previous reports on non-gynecologic malignancies. Further research is needed to find a strategy to improve clinical outcomes with ICI in this population. Objectives: Antibiotic use or BMI has been reported to be associated with the effectiveness of ICI in solid tumors. We evaluated associations between antibiotic use either prior to ICI or during ICI, BMI at ICI initiation, and oncological outcomes. Methods: We examined patients with recurrent gynecologic malignancies who underwent PD-1 monotherapy at two academic institutions. Clinical data, including antibiotic use within 30 days of initiation of ICI or during ICI treatment, BMI, tumor site, chemotherapy-free interval, prior history of radiotherapy, the objective response at 12 weeks, PFS, and OS, were assessed. Results: A total of 204 patients were eligible, of whom 22.1% (45/204) and 41.2% (84/204) received antibiotics before and during ICI, respectively. Among the patients, 18.1% (37/204) were underweight. Based on tumor site, ovarian cancer was most common (51.0%, 104/204), followed by the cervix (24.5%, 50/204) and endometrial cancer (17.6%, 36/204). About 45.1% of patients had one or two previous lines of chemotherapy, and 80.4% (164/204) patients had chemotherapy-free intervals of less than six months. In terms of overall survival, endometrial cancer showed the best survivals among gynecologic malignancies (p = 0.034). In addition, low BMI, antibiotic use before or during ICI, and chemotherapy-free interval were associated with poor overall survival. In multivariate analyses, the negative association between low BMI (HR: 2.209; 95% CI: 1.236-3.949; p=0.007) and antibiotic use during ICI (HR: 1.910; 95% CI: 1.106-3.297; p=0.02) remained significant for OS. No significant association between objective response rates at 12 weeks and BMI or antibiotics use was observed. Conclusions: Antibiotics use during ICI, or low BMI were associated with reduced survival from ICI in recurrent gynecologic malignancies, which is in line with previous reports on non-gynecologic malignancies. Further research is needed to find a strategy to improve clinical outcomes with ICI in this population.

Current antiangiogenesis therapy relies on inhibiting newly developed immature tumor blood vessels and starving tumor cells. This strategy has shown transient and modest efficacy. Here, we report a better approach to target cancer-associated endothelial cells (ECs), reverse permeability and leakiness of tumor blood vessels, and improve delivery of chemotherapeutic agents to the tumor. First, we identified deregulated microRNAs (miRs) from patient-derived cancer-associated ECs. Silencing these miRs led to decreased vascular permeability and increased maturation of blood vessels. Next, we screened a thioaptamer (TA) library to identify TAs selective for tumor-associated ECs. An annexin A2-targeted TA was identified and used for delivery of miR106b-5p and miR30c-5p inhibitors, resulting in vascular maturation and antitumor effects without inducing hypoxia. These findings could have implications for improving vascular-targeted therapy.

BVAC-C is a B cell-based and monocyte-based immuno-therapeutic vaccine transfected with a recombinant human papillomavirus (HPV) 16/18 E6/E7 gene and loaded with alpha-galactosyl ceramide, which is a natural killer T cell ligand. This phase I study sought to determine the tolerability and immunogenicity of BVAC-C in platinum-resistant recurrent cervical cancer patients. Patients with HPV 16-positive or 18-positive recurrent or persistent cervical cancer who had received at least one prior platinum-based combination chemotherapy were enrolled. BVAC-C was injected intravenously three times every four weeks, and dose escalation was planned in a three-patient cohort design at doses of 1 × 107, 4 × 107, or 1 × 108 cells/dose. Eleven patients were enrolled, and six (55%) patients had received two or more lines of platinum-based chemotherapy prior to enrollment. Treatment-related adverse events (TRAEs) were observed in 21 cycles. Most TRAEs were mild fever (n = 6.55%) or myalgia (n = 4.36%). No dose-limiting toxicities occurred. The overall response rate was 11% among nine patients evaluable, and the duration of response was 10 months. Five patients (56%) achieved a stable disease for 4.2–11 months as their best overall response. The median progression-free survival in all patients was 6.8 months (95% CI, 3.2 to infinite months), and the overall survival rate at 6 and 12 months was 89% (95% CI, 71 to 100%) and 65% (95% CI, 39 to 100%), respectively. BVAC-C induced the activation of natural killer T cells, natural killer cells, and HPV 16/18 E6/E7-specific T cells upon vaccination in all patients evaluated. BVAC-C was well tolerated and demonstrated a durable anti-tumor activity with an immune response in HPV 16-positive or 18-positive recurrent cervical carcinoma patients. A Phase 2 efficacy trial is currently underway.

Objective: This study aimed to determine risk factors associated with the failure of uterine artery ligation at its origin following development of the retroperitoneal space (UALr) and evaluated its efficacy in decreasing estimated blood loss (EBL) during single-port total laparoscopic hysterectomy (SP-TLH). Materials and methods: This study includes patient data collected prospectively from May 1st, 2013 to establish a registry for single-port surgery. Data for the present study were collected retrospectively from May 1st, 2013 to August 30th, 2016. Patients who underwent SP-TLH for a symptomatic benign disease. When bilateral UALr was performed successfully, the case was classified as part of the UALr success group. When only unilateral UALr was completed or UALr failed, the case was classified as part of the failure group. We compared patients' baseline characteristics and surgical outcomes between the two groups. Results: Bilateral UALr was successfully performed in 155 cases and failed in 64 patients. Body Mass Index (BMI) was significantly different between the two groups (24.1 kg/m(2) vs. 22.86 kg/m(2), p = 0.025). A BMI higher than 23.6 kg/m(2) was a risk factor for UALr failure in a multivariate analysis (odds ratio = 2.42, p = 0.004). EBL was significantly lower in the UALr success group compared to the UALr failure group (100 [100.0-200.0] vs. 200 [100.0-250.0], p < 0.001), and incidence of Hb decrease of more than 2 g/dl was higher in the UALr failure group (36.1% vs. 54.7%, p = 0.017). Conclusion: We identified higher BMI as a risk factor for UALr failure and demonstrated the safety and efficacy of UALr in reducing blood loss during SP-TLH. (C) 2020 Taiwan Association of Obstetrics & Gynecology. Publishing services by Elsevier B.V.

Paxillin (PXN), a key component of the focal adhesion complex, has been associated with cancer progression, but the underlying mechanisms are poorly understood. The purpose of this study was to elucidate mechanisms by which PXN affects cancer growth and progression, which we addressed using cancer patient data, cell lines, and orthotopic mouse models. We demonstrated a previously unrecognized mechanism whereby nuclear PXN enhances angiogenesis by transcriptionally regulating SRC expression. SRC, in turn, increases PLAT expression through NF-ĸB activation; PLAT promotes angiogenesis via LRP1 in endothelial cells. PXN silencing in ovarian cancer mouse models reduced angiogenesis, tumor growth, and metastasis. These findings provide a new understanding of the role of PXN in regulating tumor angiogenesis and growth.

"Metastatic malignant pheochromocytomas mimicking pelvic leiomyosarcomas: a case report." Journal of Obstetrics and Gynaecology, 41(5), pp. 838–839

Objective: The effectiveness of paclitaxel-cisplatin-ifosfamide triplet regimen (TIP) was reported to be superior to that of paclitaxel-cisplatin doublet. However, the efficacy of paclitaxel-cisplatin-bevacizumab triplet (TPA) and TIP has not been compared. Here, we compared the efficacy and safety of TIP and TPA in patients with metastatic, recurrent, or persistent cervical cancer. Methods: We retrospectively reviewed the medical records of patients with recurrent, persistent, or metastatic cervical cancer who were at the Samsung Medical Center, Seoul, Korea between January 2005 and September 2018. Of the 161 patients included in the study, 92 had received TIP and 71 had received TPA. For the study, we compared the response rates, progression-free survival (PFS), overall survival (OS), and safety in the 2 treatment groups. Results: The response rates of patients who received TIP and TPA were comparable (64.1% vs 73.2%, P = 0.239). Histology (squamous vs nonsquamous) was the only prognostic factor that affected the response to therapy (odds ratio, 0.259; 95% confidence interval [CI], 0.119-0.562; P = 0.001). The PFS after TIP and TPA treatment was similar: 12.0 months (95%CI, 10.26-13.74) vs 11.5 months (95%CI, 10.18-12.83), respectively. In a Cox proportional hazard model, objective response to therapies was the only independent prognostic factor for both PFS and OS. However, different types of therapy (TIP vs TPA) did not affect the oncological outcomes for either PFS or OS. Although hematologic toxicity was significantly higher in the TIP-treated group than in the TPA-treated group, both regimens were safe and well-tolerated. Conclusions: The effectiveness and safety of TIP was comparable to TPA in terms of response rates, survival, and adverse effects. TIP could be an effective alternative in the treatment of cervical cancer when TPA is contraindicated or unaffordable. (C) 2020 Elsevier Inc. All rights reserved.

Objectives: Adiponectin is a cytokine secreted from adipose tissue that regulates energy homeostasis, inflammation, and cell proliferation. Obesity is associated with increased risk of various cancers, including ovarian cancer. Adipokines, including adiponectin, have been implicated as a factor linking obesity and carcinogenesis. The oncogenic role of adiponectin is not known with regard to various cancer types. We sought to determine the role of adiponectin in angiogenesis in ovarian cancer in vitro. Methods: We transfected SKOV3 cells with vascular endothelial growth factor small interfering RNA in order to identify the independent angiogenic role of adiponectin in ovarian cancer. The vascular endothelial growth factor knockdown SKOV3 cell lines were treated with adiponectin for 48 h. The cytokines involved in adiponectin-mediated angiogenesis were explored using the human angiogenesis cytokine array and were verified with the enzyme-linked immunosorbent assay. The angiogenic effect of adiponectin was evaluated using the human umbilical vein endothelial cell tube formation assay. We also investigated the effects of adiponectin treatment on the migration and invasion of SKOV3 cells. Results: The number of tubes formed by human umbilical vein endothelial cell decreased significantly after knockdown of vascular endothelial growth factor (via transfection of vascular endothelial growth factor small interfering RNA into SKOV3 cells). When these vascular endothelial growth factor knockdown SKOV3 cells were treated with adiponectin, there was an increase in the number of tubes in a tube formation assay. Following adiponectin treatment, the CXC chemokine ligand 1 secretion increased in a cytokine array. This was confirmed by both enzyme-linked immunosorbent assay and Western blot. The increased secretion of CXC chemokine ligand 1 by adiponectin occurred regardless of vascular endothelial growth factor knockdown. In addition, the induction of migration and invasion of SKOV3 cells were significantly stronger with adiponectin treatment than they were without. Conclusion: Adiponectin treatment of ovarian cancer cells induces angiogenesis via CXC chemokine ligand 1 independently of vascular endothelial growth factor. These findings suggest that adiponectin may serve as a novel therapeutic target for ovarian cancer.

Antiangiogenic treatment targeting the vascular endothelial growth factor (VEGF) pathway is a powerful tool to combat tumor growth and progression; however, drug resistance frequently emerges. We identify CD5L (CD5 antigen-like precursor) as an important gene upregulated in response to antiangiogenic therapy leading to the emergence of adaptive resistance. By using both an RNA-aptamer and a monoclonal antibody targeting CD5L, we are able to abate the pro-angiogenic effects of CD5L overexpression in both in vitro and in vivo settings. In addition, we find that increased expression of vascular CD5L in cancer patients is associated with bevacizumab resistance and worse overall survival. These findings implicate CD5L as an important factor in adaptive resistance to antiangiogenic therapy and suggest that modalities to target CD5L have potentially important clinical utility.