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We report the impact of the COVID-19 pandemic on urological surgeries and hospital policies at two hospitals in Japan and Taiwan.We retrospectively surveyed the number of surgeries every 3 months in the Urology Department of Kobe University Hospital (KUH), Kobe, Japan before (January 2019-March 2020) and after (April 2020-September 2021) the COVID-19 outbreak, and in the Urology Department of Shuang Ho Hospital, Taipei Medical University (SHH-TMU), Taiwan before (January 2021-March 2021) and after (April 2021-September 2021) the outbreak, and compared the averages and types of surgery.In Kobe, COVID-19 patients were stratified such that other regional hospitals gave priority to treating COVID-19 while KUH gave priority to treating non-COVID-19 patients. In KUH, the number of surgeries did not change significantly, 237.2 ± 29.6 versus 246.3 ± 20.8 (p = 0.453). In Taiwan COVID-19 patients increased sharply in May 2021, and teaching hospitals in Taiwan were obliged to provide 20% of their total beds for COVID-19 patients. At SHH-TMU, there was a 33.3% drop in the number of surgeries during April-June 2021 compared to the pre-pandemic average. However, no significant changes were observed, 423.4 ± 68.4 versus 373 ± 91.0 (p = 0.298), because of the subsequent success in controlling the COVID-19 infection.The comparison of infection control measures between the two countries revealed that while both KUH and SHH-TMU successfully maintained the number of surgeries, the reasons for this were different for each.

Most cases of hepatocellular carcinoma (HCC) arise with the fibrotic microenvironment where hepatic stellate cells (HSCs) and carcinoma-associated fibroblasts (CAFs) are critical components in HCC progression. Therefore, CAF normalization could be a feasible therapy for HCC. Galectin-1 (Gal-1), a β-galactoside-binding lectin, is critical for HSC activation and liver fibrosis. However, few studies has evaluated the pathological role of Gal-1 in HCC stroma and its role in hepatic CAF is unclear. Here we showed that Gal-1 mainly expressed in HCC stroma, but not cancer cells. High expression of Gal-1 is correlated with CAF markers and poor prognoses of HCC patients. In co-culture systems, targeting Gal-1 in CAFs or HSCs, using small hairpin (sh)RNAs or an therapeutic inhibitor (LLS30), downregulated plasminogen activator inhibitor-2 (PAI-2) production which suppressed cancer stem-like cell properties and invasion ability of HCC in a paracrine manner. The Gal-1-targeting effect was mediated by increased a disintegrin and metalloprotease 17 (ADAM17)-dependent TNF-receptor 1 (TNFR1) shedding/cleavage which inhibited the TNF-α → JNK → c-Jun/ATF2 signaling axis of pro-inflammatory gene transcription. Silencing Gal-1 in CAFs inhibited CAF-augmented HCC progression and reprogrammed the CAF-mediated inflammatory responses in a co-injection xenograft model. Taken together, the findings uncover a crucial role of Gal-1 in CAFs that orchestrates an inflammatory CSC niche supporting HCC progression and demonstrate that targeting Gal-1 could be a potential therapy for fibrosis-related HCC.

Biochemical parameters in the serum of mice before and after treatment with LCC03 (40 mg/kg) for 5 weeks.

Metastatic castration-resistant prostate cancer (CRPC) is currently incurable. Cancer growth and progression is intimately affected by its interaction with host microenvironment. Cotargeting of the stroma and prostate cancer is therefore an emerging therapeutic strategy for metastatic CRPC. Cancer-induced osteoclastogenesis is known to contribute to CRPC bone metastasis. This study is to extend pharmacologic value of our synthesized LCC03, a derivative of 5-(2′,4′-difluorophenyl)-salicylanilide that has previously testified for its osteoclastogenesis activity, by exploring its additional cytotoxic properties and underlying mechanism in CRPC cells. LCC03 was chemically synthesized and examined for cell growth inhibition in a serial of CRPC cell lines. We demonstrated that LCC03 dose-dependently suppressed proliferation and retarded cell-cycle progression in CRPC cells. The classical autophagy features, including autophagosome formation and LC3-II conversion, were dramatically shown in LCC03-treated CRPC cells, and it was associated with the suppressed AKT/mTOR signaling pathways, a major negative regulator of autophagy. Moreover, an expanded morphology of the endoplasmic reticulum (ER), increased expression of the ER stress markers GRP78 and PERK, and eIF2α phosphorylation were observed. Blockage of autophagy and PERK pathways using small molecule inhibitors or shRNA knockdown reversed LCC03-induced autophagy and cell death, thus indicating that the PERK–eIF2α pathway contributed to the LCC03-induced autophagy. Furthermore, treatment of tumor-bearing mice with intraperitoneal administered LCC03 suppressed the growth of CRPC xenografts in mouse bone without systemic toxicity. The dual action of 5-(2′,4′-difluorophenyl)-salicylanilide on targeting both the osteoclasts and the tumor cells strongly indicates that LCC03 is a promising anticancer candidate for preventing and treating metastatic CRPC.

Effect of mTOR activation on LCC03-induced PERK expression

Supplementary Data 1-5 from Coevolution of Prostate Cancer and Bone Stroma in Three-Dimensional Coculture: Implications for Cancer Growth and Metastasis

Supplementary Table S1. Comparison of fold change of gene expression in ADAM9 knockdown and brain-metastatic cell sublines (Bm2 and Bm7) with parental cells.

Supplementary Data 6-9 from Coevolution of Prostate Cancer and Bone Stroma in Three-Dimensional Coculture: Implications for Cancer Growth and Metastasis

Supplementary Legends 1-9 from Coevolution of Prostate Cancer and Bone Stroma in Three-Dimensional Coculture: Implications for Cancer Growth and Metastasis

The transmembrane cell adhesion protein ADAM9 has been implicated in cancer cell migration and lung cancer metastasis to the brain, but the underpinning mechanisms are unclear and clinical support has been lacking. Here, we demonstrate that ADAM9 enhances the ability of tissue plasminogen activator (tPA) to cleave and stimulate the function of the promigratory protein CDCP1 to promote lung metastasis. Blocking this mechanism of cancer cell migration prolonged survival in tumor-bearing mice and cooperated with dexamethasone and dasatinib (a dual Src/Abl kinase inhibitor) treatment to enhance cytotoxic treatment. In clinical specimens, high levels of ADAM9 and CDCP1 correlated with poor prognosis and high risk of mortality in patients with lung cancer. Moreover, ADAM9 levels in brain metastases derived from lung tumors were relatively higher than the levels observed in primary lung tumors. Our results show how ADAM9 regulates lung cancer metastasis to the brain by facilitating the tPA-mediated cleavage of CDCP1, with potential implications to target this network as a strategy to prevent or treat brain metastatic disease. Cancer Res; 74(18); 5229–43. ©2014 AACR.

Medulloblastoma (MB) was classified into four molecular subgroups: WNT, SHH, group 3, and group 4. In 2017, 12 subtypes within 4 subgroups and 8 subtypes within non-WNT/non-SHH subgroups according to the differences of clinical features and biology were announced. In this study, we aimed to identify the heterogeneity of molecular features for discovering subtype specific factors linked to diagnosis and prognosis. We retrieved 70 MBs in children to perform RNA sequencing and a DNA methylation array in Taiwan. Integrated with clinical annotations, we achieved classification of 12 subtypes of pediatric MBs in our cohort series with reference to the other reported series. We analyzed the correlation of cell type enrichment in SHH MBs and found that M2 macrophages were enriched in SHH β, which related to good outcomes of SHH MBs. The high infiltration of M2 macrophages may be an indicator of a favorable prognosis and therapeutic target for SHH MBs. Furthermore, C11orf95-RELA fusion was observed to be associated with recurrence and a poor prognosis. These results will contribute to the establishment of a molecular diagnosis linked to prognostic indicators of relevance and help to promote molecular-based risk stratified treatment for MBs in children.

AXL which is a chemosensitizer protein for breast cancer cells in response to epidermal growth factor receptor-tyrosine kinase inhibitor and suppresses tumor growth. The clinical information show nuclear factor I (NFI)-C and NFI-X expression correlate with AXL expression in breast cancer patients. Following, we establish serial deletions of AXL promoter to identify regions required for Adenovirus-5 early region 1A (E1A)-mediated AXL suppression. All of the NFI family members were extensively studied for their expression and functions in regulating AXL. Moreover, E1A post-transcriptionally downregulates AXL expression through NFI. NFI-C and NFI-X, not NFI-A and NFI-B, resulting in cell death in response to EGFR-TKI. Our finding suggests that NFI-C and NFI-X are crucial regulators for AXL and significantly correlated with poor survival of breast cancer patients.

BACKGROUND Focal therapies for prostate cancer (PC) can reduce adverse events and do not lead to androgen-independent progression. Ultrasound could be used for cancer treatments if the repetition frequency is fitted to the purpose. We investigated the possible therapeutic effect of ultrasound irradiation on PC cells. MATERIALS AND METHODS We irradiated two PC cell lines, androgen-dependent LNCaP and -independent PC-3 with ultrasound (3.0 W/cm2 , 3 MHz, irradiation time rate: 20%) for 2 minutes for 1 day or 3 consecutive days at a repetition frequency of 1, 10, or 100 Hz in vitro. Cell proliferation and apoptosis were determined after irradiation. RESULTS Cell proliferation of PC-3 was significantly inhibited after 1 day (P

Distant metastasis is the leading cause of death in patients with breast cancer. Despite considerable treatment advances, the clinical outcomes of patients with metastatic breast cancer remain poor. CSCs can self-renew, enhancing cancer progression and metastasis. Dicer, a microRNA (miRNA) processing-related enzyme, is required for miRNA maturation. Imbalanced Dicer expression may be pivotal in cancer progression. However, whether and how Dicer affects the stemness of metastatic breast cancer cells remains unclear. Here, we hypothesized that Dicer regulates the migration, invasion, and stemness of breast cancer cells. We established highly invasive cell lines (MCF-7/I-3 and MDA-MB-231/I-3) and observed that Dicer expression was conspicuously lower in the highly invasive cells than in the parental cells. The silencing of Dicer significantly enhanced the cell migratory/invasive abilities and CSCs properties of the breast cancer cells. Conversely, the overexpression of Dicer in the highly invasive cells reduced their migration, invasion, and CSCs properties. Our bioinformatics analyses demonstrated that low Dicer levels were correlated with increased breast cancer risk. Suppression of Dicer inhibited miR-200b expression, whereas miR-200b suppression recovered Dicer knockdown-induced migration, invasion, and cancer stem cells (CSCs) properties of the breast cancer cells. Thus, our findings reveal that Dicer is a crucial regulator of the migration, invasion, and CSCs properties of breast cancer cells and is significantly associated with poor survival in patients with breast cancer.

The high metastasis and drug resistance of colorectal cancer (CRC) cause high mortality and rank third among the top ten cancer deaths. TY1 is a biflavonoid isolated from Taxus x media cv. Hicksii. We have previously reported that TY1 inhibits metastatic behavior by inhibiting F-actin polymerization. However, the biological activities of TY1 to overcome drug resistance remain unclear. In this study, we investigated the pharmacological mechanism of TY1 for inhibiting 5-fluorouracil (5-FU)-resistant CRC cells and its therapeutic effect. The 5-FU sensitive (COLO205) and 5-FU resistant (HT-29) CRC cells were used to determine the antitumor activity of TY1. Bioinformatic analysis was conducted to explore the antitumor effects of TY1 in 5-FU resistant HT-29 cells. The effects of TY1 on 5-FU-resistant CRC cells were determined by cell-based analyses, including MTT assay, cellular and mitochondrial ROS-superoxide detection, and confocal images. Finally, the therapeutic effects of TY1 were evaluated in HT-29 xenograft CRC mice. We found TY1 inhibited cell proliferation but did not affect the cell cycle. TEM images showed that TY1 induced vacuole production. Transcriptome data indicated that interferon-related genes specifically mitochondrial interferon alpha-inducible protein (IFIP) were significantly decreased after TY1 treatment. TY1 increased oxidative stress generated by reactive oxygen species (ROS) and superoxide. When IFIP was overexpressed, TY1-induced oxidative stress was reduced. In xenograft CRC mouse, TY1 effectively inhibited tumor growth. The anticancer activity of TY1 is attributed to TY1-induced ROS and superoxide by downregulating mitochondrial IFIP expression. Therefore, these data suggest that TY1 may be a promising new compound in the treatment of 5-FU-resistant CRC. Citation Format: Ting-Yan Jian, Shian-Ying Sung, Yu-Ling Lin. A biflavonoid TY1 induced mitochondrial oxidative stress in 5-fluorouracil-resistant colorectal cancer cells to suppress tumor progression. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4782.

Introduction: Medulloblastoma (MB) is the most common malignant tumor in children. Since most MB-related deaths are the result of metastasis, current standard care for MB is aggressive with intense post-surgical radiotherapies, which aim to minimize metastatic occurences and are applied in most cases regardless of tumor subgroups. Among the four molecular subgroups, sonic-hedgehog (SHH-) MB has a high prevalence and well-characterized profiles. A proportion of SHH-MB cases with higher metastasis rate and worse prognosis have been linked to aberrant expressions of certain genes such as MYCN. Strategies to target such pro-metastatic drivers have been explored but proven to be challenging. Therefore, discovery of novel targets in MB is an urgent need to optimize treatment and improve outcomes. Long non-coding RNAs (lncRNAs) have recently emerged as malignancy regulators in many cancers. More insights into the role of lncRNAs in metastasis-promoting phenotypes, especially cancer stemness, would present both prognostic and therapeutic significance in metastatic MB. Objective: To determine the role of lncRNA LOXL1-AS1 in progression and metastasis of SHH-MB. Methods: First, we used RNA-sequencing to screen for differentially expressed lncRNAs in SHH-MB cells (Daoy) achieved a higher malignancy level via MYCN overexpression (Daoy-MYCN), identifying LOXL1-AS1 as one of the pro-malignant lncRNA candidates. LOXL1-AS1 expressions and prognostic associations were validated in clinical samples and datasets. Next, MB cells with stable LOXL1-AS1 knockdown (Daoy-MYCN) or overexpression (Daoy and ONS-76) were subjected to migration and invasion assays, as well as seeded in sphere-forming condition to assess sphere formation capacities and stemness marker expressions. Finally, we injected Daoy-MYCN cells knockdown of LOXL1-AS1 into BALB/c mouse brain to observe formation of tumor, metastasis, and survival rates. Results: LOXL1-AS1 was increased in Daoy cells under MYCN overexpression, was highly expressed in MB compared to normal brain, and was associated with poor prognosis in SHH-MB, particularly in the SHH-α and SHH-γ subgroups. MB cells with LOXL1-AS1 knockdown or overexpression showed a significant decrease or increase, respectively, in cell migration, cell invasion, sphere numbers, sphere diameters, and expression of stemness markers. Mice orthotopically transplanted with LOXL1-AS1-knockdown cells experienced a delayed tumor growth and metastasis development with improved overall and metastasis-free survivals. Conclusion: Our study has proven the role of LOXL1-AS1 in pro-metastatic phenotypes of MYCN-associated SHH-MB through a possible mechanism of inducing cancer stemness, which expands our current array of lncRNA candidates to develop MB-targeted therapies. The mechanisms of LOXL1-AS1-driven metastasis are being investigated in MB models. Citation Format: Anh Duy Do, Chia-Ling Hsieh, Tai-Tong Wong, Shian-Ying Sung. The pro-metastatic role of long-noncoding RNA LOXL1-AS1 in sonic-hedgehog medulloblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1285.

Klebsiella pneumoniae is a typical pathogen in urinary tract infections (UTI), and the emergence of extended spectrum beta-lactamase (ESBL)-producing strains has been frequently reported, accompanied by higher quinolone resistance rates. There are two major mechanisms of quinolone resistance, mutations in quinolone resistance-determining regions (QRDR) and the presence of the plasmid-mediated quinolone resistance (PMQR) genes. This study aimed to investigate quinolone resistance among 105 ESBL-producing K. pneumoniae specimens isolated from UTI patients in Indonesia. These were characterized for antimicrobial resistance to nalidixic acid, ciprofloxacin, and levofloxacin, QRDR mutations in gyrA and parC and the presence of PMQR genes. We found that 84.8% of the collected isolates were resistant to at least one of the quinolones. QRDR mutation in gyrA was observed in 49.5% of these strains and parC mutations in 61.0%. PMQR genes were identified in 84.8% of strains. The QRDR mutations clearly had a greater effect on resistance than the PMQR genes. In conclusion, we found high quinolone resistance rates in Indonesian ESBL-producing K. pneumoniae, in which QRDR mutation played a major role.

Background Hypermucoviscous (HMV) Klebsiella pneumoniae produces large amounts of capsular polysaccharides, leading to high mortality. Since extended spectrum beta-lactamase (ESBL)-producing HMV K. pneumoniae strains have increased in Japan, we investigated and compared the antimicrobial susceptibilities and genetic characteristics of HMV and non-HMV ESBL-producing K. pneumoniae. Methods We investigated 291 ESBL-producing K. pneumoniae collected between 2012 and 2018, and in them 54 HMV strains were identified and comparable 53 non-HMV strains were selected. Then, ESBL gene detection, plasmid replicon typing, and virulence gene detection were done by PCR amplification. Results Almost all of the HMV K. pneumoniae strains possessed uge (98.1%), wabG (96.3%), rmpA (94.4%), iucA (79.6%), fimH (70.4%), iroB (70.4%), and peg-344 (70.4%). These genes were found less frequently in non-HMV strains (uge 20.8%, wabG 83.0%, rmpA 7.5%, iucA 3.8%, fimH 9.4%, iroB 5.7%, and peg-344 1.9%). K2 capsule type (40.7%) was most common in HMV strains. HMV strains showed higher resistance to cefepime (p = 0.001) and piperacillin/tazobactam (p = 0.005) than non-HMV strains. CTX-M-15 (75.9%, 60.4%) was the dominant ESBL type in both HMV and non-HMV strains, and the most common plasmid replicon type was IncFII (52.1%) in CTX-M-15-producing strains. Conclusions We found that HMV strains had more virulence genes and showed higher resistance to antibiotics than non-HMV strains. The most common capsule type was K2. CTX-M-15 was the most common type of ESBL gene in both HMV and non-HMV strains in Japan. The FII plasmid might be related to the spread of CTX-M-15 among K. pneumoniae strains.

Treating brain tumors presents enormous challenges, and there are still poor prognoses in both adults and children. Application of novel targets and potential drugs is hindered by the function of the blood-brain barrier, which significantly restricts therapeutic access to the tumor. Mesenchymal stem cells (MSCs) can cross biological barriers, migrate to sites of injuries to exert many healing effects, and be engineered to incorporate different types of cargo, making them an ideal vehicle to transport anti-tumor agents to the central nervous system. Extracellular vesicles (EVs) produced by MSCs (MSC-EVs) have valuable innate properties from parent cells, and are being exploited as cell-free treatments for many neurological diseases. Compared to using MSCs, targeted delivery via MSC-EVs has a better pharmacokinetic profile, yet avoids many critical issues of cell-based systems. As the field of MSC therapeutic applications is quickly expanding, this article aims to give an overall picture for one direction of EV-based targeting of brain tumors, with updates on available techniques, outcomes of experimental models, and critical challenges of this concept.

The development of novel magnetic nanoparticles (MNPs) with satisfactory biocompatibility for biomedical applications has been the subject of extensive exploration over the past two decades. In this work, we synthesized superparamagnetic iron oxide MNPs coated with polystyrene sulfonic acid (PSS-MNPs) and with a conventional co-precipitation method. The core size and hydrodynamic diameter of the PSS-MNPs were determined as 8–18 nm and 50–200 nm with a transmission electron microscopy and dynamic light scattering, respectively. The saturation magnetization of the particles was measured as 60 emu g−1 with a superconducting quantum-interference-device magnetometer. The PSS content in the PSS-MNPs was 17% of the entire PSS-MNPs according to thermogravimetric analysis. Fourier-transform infrared spectra were recorded to detect the presence of SO3− groups, which confirmed a successful PSS coating. The structural properties of the PSS-MNPs, including the crystalline lattice, composition and phases, were characterized with an X-ray powder diffractometer and 3D nanometer-scale Raman microspectrometer. MTT assay and Prussian-blue staining showed that, although PSS-MNPs caused no cytotoxicity in both NIH-3T3 mouse fibroblasts and SK-HEP1 human liver-cancer cells up to 1000 μg mL−1, SK-HEP1 cells exhibited significantly greater uptake of PSS-MNPs than NIH-3T3 cells. The low cytotoxicity and high biocompatibility of PSS-MNPs in human cancer cells demonstrated in the present work might have prospective applications for drug delivery., GRAPHICAL ABSTRACT

Purpose: The aim of this study was to compare and investigate the efficacy of using the 5α-reductase inhibitor dutasteride after holmium laser enucleation of the prostate (HoLEP) to improve postoperative urination and surgery-related complications. Methods: This is a retrospective observational study comparing patients who received or did not receive 5α-reductase inhibitors prior to HoLEP. Of a total of 270 patients, 40 received the 5α-reductase inhibitor dutasteride. We compared the factors including age, postoperative maximal flow rate (MFR; mL/s), postoperative prostate-specific antigen (PSA) (ng/mL), preoperative MFR (mL/s), preoperative PSA (ng/mL), prostate cancer (%), operative time (min), preoperative residual urine (mL), postoperative residual urine (mL), urinary incontinence (day 1; %), urinary incontinence (1 month; %), urinary incontinence (3 months; %), urethral catheter indwelling period (days), morcellation time (min), enucleation time (min), intraoperative complications (%), postoperative complications (%), prostate volume (mL), enucleated weight (g), and hospitalization period (days). Results: Postoperative PSA (p = 0.0071), morcellation time (p = 0.0444), postoperative complications (p = 0.0350) and prostate volume (p = 0.0069), but not enucleated prostate weight (p = 0.8809), were significantly lower in the dutasteride group. Importantly, enucleation efficiency and morcellation efficiency did not show any significant difference between the dutasteride and the non-dutasteride groups. Conclusions: Use of a preoperative 5α-reductase inhibitor significantly correlated with surgery-related factors, with less morcellation time, fewer postoperative complications, and lower postoperative PSA. Surgeons performing HoLEP may wish to take these findings into account.

To investigate the role of sonic hedgehog (Shh) signaling and epithelial–mesenchymal transition (EMT) in bladder cancer progression and invasion. We cultured three bladder cancer cell lines, muscle-invasive T24 and 5637, and non-muscle-invasive KK47, in the presence of a recombinant-Shh (r-Shh) protein or cyclopamine, a Shh signaling inhibitor, to investigate proliferation and expression of EMT markers. Wound-healing assays and transwell assay were performed to evaluate cell invasion and migration. Mice were then inoculated with bladder cancer cells and treated with cyclopamine. Mouse tumor samples were stained for Shh signaling and EMT markers. R-Shh protein enhanced cell proliferation, whereas cyclopamine significantly suppressed cell proliferation, especially in invasive cancer (5637 and T24) (p

Introduction: Medulloblastoma (MB) is the most common malignant tumor in children. MB-related treatment failures and deaths mostly result from metastasis. One of the key genetic drivers in MB is MYCN, whose aberrant expressions are associated with tumor aggressiveness and poor prognosis. Long non-coding (lnc)RNAs have been implied as potential targets for cancer treatment. More insights into the role of lncRNAs in MYCN-mediated MB progression would have both prognostic and therapeutic significance in metastatic MB. Cell-derived vesicles such as exosome can encapsulate siRNAs to improve their bioavailability and targeted delivery to brain tumors. Mesenchymal stem cell (MSC) is a readily available and modifiable source to produce functional exosomes. Objective: To identify the downstream lncRNAs of MYCN that contributes to MB metastasis, and to develop a novel gene therapy for MB by MSC exosome-delivered siRNAs to target these prometastatic lncRNAs. Methods: An RNA sequencing analysis was used to identify LOXL1-AS1 as a MYCN targeting gene in MYCN-overexpressing MB cells. The association of LOXL1-AS1 and MYCN was validated in MB cell lines by RT-qPCR and Western blot assay, and in tumor tissues by in-situ hybridization and immunohistochemistry. The anti-tumor effect of LOXL1-AS1 knockdown in MYCN-overexpressing MB cells was assessed in vitro by migration and sphere formation assays, and in an orthotopic xenograft mouse model by intracranial injection. To generate therapeutic exosomes, MSCs were transfected with exogenous LOXL1-AS1 siRNAs and then subjected to exosome characterization of size distribution and marker expression. The effect of exosome treatment on MB cells was evaluated using RT-qPCR and wound healing assay. Results: Elevated LOXL1-AS1 was found in MYCN-overexpressed MB cells compared to parental cells, and this association was confirmed in clinical tissue samples. LOXL1-AS1 expression was correlated with poor prognosis in SHH-α and SHH-γ MB subtypes of patients. Knockdown of LOXL1-AS1 significantly reduced cell migration and cancer stemness properties of MB cells in vitro, and suppressed orthotopic tumor growth, metastasis, and enhanced survival in xenografted mice. Exogenous siRNAs were successfully packaged into MSC exosomes by cell transfection. Treatment of MB cells with LOXL1-AS1 siRNA-loaded exosomes effectively reduced LOXL1-AS1 expression and inhibited cell migration compared to those treated with unmodified exosomes. Conclusion: Our study has proven the contributive role of LOXL1-AS1 in promoting cancer progression and metastasis of MYCN-associated MB, and provided in vitro evidence to support the feasibility of LOXL1-AS1 gene therapy through siRNA-loaded MSC exosomes. In vivo biodistribution and therapeutic efficacy assessment of MSC exosome-delivered LOXL1-AS1 siRNAs are being investigated in animal models. Citation Format: Anh Duy Do, Chia-Ling Hsieh, Tai-Tong Wong, Shian-Ying Sung. LOXL1-AS1 contributes to medulloblastoma metastasis: A potential target for mesenchymal stem cell exosome-delivered gene therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5369.

Introduction: Cancer-associated fibroblasts (CAFs), the main cellular component of the tumor microenvironment (TME), contribute to tumor metastasis. Prostate cancer (PCa) is the most common malignancy among the elderly male population, and it is typically fatal when the disease progresses to metastasis and castration-resistant stage (CRPC). We have previously demonstrated that ADAM9 increased during PCa progression, and ROS is the critical mediator regulating ADAM9 expression in CRPC cells to promote tumor growth. In addition to membrane-anchored ADAM9, a secreted variant of ADAM9 (sADAM9) was recently identified in activated liver stromal cells that facilitated colon cancer liver metastasis. However, it is not clear whether sADAM9 is expressed by prostatic CAFs and, if so, whether it contributes to aggressive tumor phenotypes through tumor-stromal interactions. Objective: To investigate the sADAM9 variants in CAFs and to elucidate the effects of CAF-derived sADAM9 on the aggressiveness of PCa cells. Methods: Immunohistochemistry staining and ELISA of clinical samples were used to assess the expression of ADAM9 during PCa tumor progression. Laser microdissected tissues, primary cell lines, and experimental cell model of paired normal fibroblasts and CAF cells were analyzed. The effects of sADAM9 on CRPC progression were conducted in PC3 cells by WST-1 cell proliferation assay, transwell migration assay, and μ-slide chemotaxis assay. Immunoblot analyses of integrin β1 and migration-related kinases were used to verify the possible downstream signaling pathway of sADAM9. Results: Increased overall intensity of ADAM9 protein expression in PCa patients with metastasis compared. Elevated ADAM9 expression was observed in both cancerous epithelial cells and the tumor stroma. The average serum ADAM9 concentration measured by ELISA was 6.253 ± 0.448 ng/mL in BPH patient samples versus 10.89 ± 0.953 ng/mL in PCa patient samples. A smaller molecular weight (~50-kDa) of ADAM9 was detected in CM from cultured CAFs but not normal fibroblasts or PCa cell lines except for DU145 cells. Treatment of PC3 cells with sADAM9 induced a significant chemotactic migration while did not affect cell proliferation. Furthermore, a stark increase in activated integrin β1, phosphorylated focal adhesion kinase (p-FAK), and p-AKT, but not p-mTOR in PCa cells by sADAM9 suggested the induction of cell migration occurs through activation of the integrin β1/FAK signaling pathways. Conclusion: Herein, we have provided in vitro evidence to demonstrate the pro-metastatic function of sADAM9 from CAFs that facilitate the migration of CRPC cells through integrin receptor signaling, which could be partly responsible for the promotion of CRPC metastasis. Further molecular identification of controlling spliced ADAM9 expression in CAFs remains to investigate. Citation Format: Weichieh Chen, Le Thi Huynh Trang, Chia-Ling Hsieh, Katsumi Shigemura, Masato Fujisawa, Leland W.K. Chung, Shian-Ying Sung. ADAM9 secreted by cancer-associated stromal cells promotes metastasis of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6125.

BACKGROUND/AIM Epithelial-mesenchymal transition (EMT) via Sonic Hedgehog (Shh) signaling may be one of the mechanisms of progression of castration-resistant prostate cancer (CRPC). In this study, we investigated the possible therapeutic effect of vismodegib, a new Shh inhibitor, in a mouse CRPC model. MATERIALS AND METHODS We determined cell proliferation, apoptosis and the expression of EMT-related genes for three prostate cancer cell lines; androgen-dependent LNCaP and independent C4-2B and PC-3 in the presence of vismodegib in vitro. Fifty mg/kg of vismodegib were orally administered into mice bearing C4-2B and PC-3 tumors, respectively every other week for 3 weeks. RESULTS Vismodegib significantly inhibited cell proliferation and induced cell apoptosis in all cell lines in vitro (p

This article describes cell signaling network of metastatic prostate cancer (PCa) to bone and visceral organs in the context of tumor microenvironment and for the development of novel therapeutics. The article focuses on our recent progress in the understanding of: 1) The plasticity and dynamics of tumor–stroma interaction; 2) The significance of epigenetic reprogramming in conferring cancer growth, invasion and metastasis; 3) New insights on altered junctional communication affecting PCa bone and brain metastases; 4) Novel strategies to overcome therapeutic resistance to hormonal antagonists and chemotherapy; 5) Genetic-based therapy to co-target tumor and bone stroma; 6) PCa-bone-immune cell interaction and TBX2-WNTprotein signaling in bone metastasis; 7) The roles of monoamine oxidase and reactive oxygen species in PCa growth and bone metastasis; and 8) Characterization of imprinting cluster of microRNA, in tumor–stroma interaction. This article provides new approaches and insights of PCa metastases with emphasis on basic science and potential for clinical translation. This article referenced the details of the various approaches and discoveries described herein in peer-reviewed publications. We dedicate this article in our fond memory of Dr. Donald S. Coffey who taught us the spirit of sharing and the importance of focusing basic science discoveries toward translational medicine. Keywords: Prostate cancer, Castration resistance, Metastasis, Cancer-stromal interaction, Targeted therapy