Prostate Cancer Invasion Research Articles

MiR-183-5p promotes migration and invasion of prostate cancer by targeting TET1

BackgroundProstate cancer (PCa) is one of the common malignant tumors worldwide. MiR-183-5p has been reported involved in the initiation of human PCa, this study aimed to investigate whether miR-183-5p affects the development of prostate cancer.MethodsIn this study, we analyzed the expression of miR-183-5p in PCa patients and its correlation with clinicopathological parameters based on TCGA data portal. CCK-8, migration assay and invasion and wound-healing assay were performed to detect proliferation, migration and invasion in PCa cells.ResultsWe found the expression of miR-183-5p was significantly increased in PCa tissues, and high expression of miR-183 was positively associated with poor prognosis of PCa patients. Over-expression of miR-183-5p promoted the migration, invasion capacities of PCa cells, whereas knockdown of miR-183-5p showed reversed function. Furthermore, luciferase reporter assay showed TET1 was identified as a direct target of miR-183-5p, which was negatively correlation with miR-183-5p expression level. Importantly, rescue experiments demonstrated TET1 over-expression could reverse miR-183-5p mimic induced-acceleration of PCa malignant progression.ConclusionOur results indicated that miR-183-5p could act as a tumor promoter in PCa and it accelerated the malignant progression of PCa by directly targeting and down-regulating TET1.

Comparison of Magnetic Resonance Imaging-Based Radiomics Features with Nomogram for Prediction of Prostate Cancer Invasion.

To explore the value of the magnetic resonance imaging (MRI) radiomics model in predicting prostate cancer (PCa) invasion. Clinical data of 86 pathologically confirmed PCa patients in our hospital were collected, including 44 cases in the invasive group and 42 cases in the non-invasive group. All patients underwent MRI examinations, and the same parameters were used. The lesion area was manually delineated and the radiomics features were extracted from T2WI. The radiomics signature based on LASSO regression was established. Besides, logistic regression was used to identify independent clinical predictors, and a combined model incorporating the radiomics signature and independent clinical risk factor was constructed. Finally, the receiver operating characteristic curve (ROC) analysis and decision curve analysis (DCA) was performed to compare the prediction efficiency and clinical benefit of each model. A total of 867 radiomics features were obtained, and six of them were incorporated into the radiomics model. Multivariate logistic regression analysis exhibited the Gleason score as an independent clinical risk factor for PCa invasion. ROC results showed that the performance of the radiomics model was comparable to that of the clinical-radiomics model in predicting PCa invasion, and it was better than that of the single Gleason score. DCA also confirmed the considerable clinical application value of the radiomics and the clinical-radiomics models. As a simple, non-invasive, and efficient method, the radiomics model has important predictive value for PCa invasion.

Reliable Visualization of the Treatment Effect of Transperineal Focal Laser Ablation in Prostate Cancer Patients by Magnetic Resonance Imaging and Contrast-enhanced Ultrasound Imaging

BackgroundTransperineal focal laser ablation (TPLA) treatment for prostate cancer (PCa) is an experimental focal ablative therapy modality with low morbidity. However, a dosimetry model for TPLA is lacking. ObjectiveTo determine (1) the three-dimensional (3D) histologically defined ablation zone of single- and multifiber TPLA treatment for PCa correlated with magnetic resonance imaging (MRI) and contrast-enhanced ultrasound (CEUS) and (2) a reliable imaging modality of ablation zone volumetry. Design, setting, and participantsThis was a prospective, multicenter, and interventional phase I/II pilot study with an ablate-and-resect design. TPLA was performed in 12 patients with localized prostate cancer divided over four treatment regimens to evaluate potential variation in outcomes. InterventionTPLA was performed approximately 4 wk prior to robot-assisted radical prostatectomy (RARP) in a daycare setting using local anesthesia. Outcome measurements and statistical analysisFour weeks after TPLA, ablation zone volumetry was determined on prostate MRI and CEUS by delineation and segmentation into 3D models and correlated with whole-mount RARP histology using the Pearson correlation index. Results and limitationsTwelve office-based TPLA procedures were performed successfully under continuous transrectal ultrasound guidance using local perineal anesthesia. No serious adverse events occurred. A qualitative analysis showed a clear demarcation of the ablation zone on T2-weighted MRI, dynamic contrast-enhanced MRI, and CEUS. On pathological evaluation, no remnant cancer was observed within the ablation zone. Ablation zone volumetry on CEUS and T2-weighted MRI compared with histology had a Pearson correlation index of r = 0.94 (95% confidence interval [CI] 0.74–0.99, p < 0.001) and r = 0.93 (95% CI 0.73–0.98, p < 0.001), respectively. ConclusionsCEUS and prostate MRI could reliably visualize TPLA ablative effects after minimally invasive PCa treatment with a high concordance with histopathological findings and showed no remnant cancer. Patient summaryThe treatment effects of a novel minimally invasive ablation therapy device can reliably be visualized with radiological examinations. These results will improve planning and performance of future procedures.

Progressive Spreading of DNA Methylation in the GSTP1 Promoter CpG Island across Transitions from Precursors to Invasive Prostate Cancer.

DNA hypermethylation at the GSTP1 promoter progressively spreads from being unmethylated in normal prostate to intermediate levels in precursor lesions to extensive methylation in cancer. This molecular progression of GSTP1 promoter methylation patterns in early prostate carcinogenesis could be useful for identification and interception of prostate cancer precursors.

Retraction Note to: miR-217-5p inhibits invasion and metastasis of prostate cancer by targeting clusterin.

Retraction Note to: miR-217-5p inhibits invasion and metastasis of prostate cancer by targeting clusterin.

Neuropeptide Y Peptide Family and Cancer: Antitumor Therapeutic Strategies.

Currently available data on the involvement of neuropeptide Y (NPY), peptide YY (PYY), and pancreatic polypeptide (PP) and their receptors (YRs) in cancer are updated. The structure and dynamics of YRs and their intracellular signaling pathways are also studied. The roles played by these peptides in 22 different cancer types are reviewed (e.g., breast cancer, colorectal cancer, Ewing sarcoma, liver cancer, melanoma, neuroblastoma, pancreatic cancer, pheochromocytoma, and prostate cancer). YRs could be used as cancer diagnostic markers and therapeutic targets. A high Y1R expression has been correlated with lymph node metastasis, advanced stages, and perineural invasion; an increased Y5R expression with survival and tumor growth; and a high serum NPY level with relapse, metastasis, and poor survival. YRs mediate tumor cell proliferation, migration, invasion, metastasis, and angiogenesis; YR antagonists block the previous actions and promote the death of cancer cells. NPY favors tumor cell growth, migration, and metastasis and promotes angiogenesis in some tumors (e.g., breast cancer, colorectal cancer, neuroblastoma, pancreatic cancer), whereas in others it exerts an antitumor effect (e.g., cholangiocarcinoma, Ewing sarcoma, liver cancer). PYY or its fragments block tumor cell growth, migration, and invasion in breast, colorectal, esophageal, liver, pancreatic, and prostate cancer. Current data show the peptidergic system's high potential for cancer diagnosis, treatment, and support using Y2R/Y5R antagonists and NPY or PYY agonists as promising antitumor therapeutic strategies. Some important research lines to be developed in the future will also be suggested.

Abstract A017: Molecular analysis of matched PIN, invasive prostate cancer, and adjacent normal prostate tissue samples reveal distinct transcriptional signatures and clonal relationships

Abstract Background: Prostate cancer (PCa) is highly heterogeneous, and while some tumors remain indolent throughout a patient's lifetime, others are highly aggressive, leading to significant morbidity and death. The current methods for PCa diagnosis and prognosis are highly suboptimal, resulting in over-diagnosis and overtreatment of many clinically insignificant tumors. To understand the nature of PCa heterogeneity, we conducted comprehensive genomic analyses on matched samples of adjacent normal prostate tissue, prostate intraepithelial neoplasia (PIN), and invasive PCa. Methods: To investigate the changes cells undergo during PCa development and progression, we analyzed genomic and transcriptomic alterations in 92 adjacent normal, 80 PIN, and 99 PCa samples from 34 patients. For each sample type, both epithelial and stromal components were analyzed separately using laser capture microdissection of formalin fixed paraffin embedded tissue. Transcriptomic analysis was performed using smart3-SEQ and copy number analysis was performed by low-pass whole genome sequencing (WGS). The samples were analyzed as three separate data sets. Results: We studied the spectrum of molecular changes present in both epithelial and stromal samples. A negative-binomial regression model was used to identify differentially expressed genes between adjacent normal prostate tissue and invasive PCa in epithelium and stroma separately using the discovery data set. The resulting gene lists were used to perform non-negative matrix factorization clustering of all samples in the three datasets separately. This analysis identified two epithelial and two stromal clusters with distinct RNA expression profiles in all three data sets. For invasive PCa samples, all except three epithelial samples and five stromal samples clustered in the ‘invasive-like’ clusters. Similarly for adjacent normal samples, all except three of the epithelial and six of the stromal samples clustered as ‘normal-like’ across the three datasets. For PIN samples, 54% of epithelial and 62% of stromal samples classified as ‘normal-like’. We performed phylogenetic analysis of WGS data and identified PIN lesions with and without clear clonal relation to invasive PCa lesions. Recurrent copy number variations were identified in both PIN and PCa lesions. Conclusion. We identified two distinct epithelial and two distinct stromal expression clusters with ‘invasive-like’ and ‘normal-like’ signatures. WGS analysis revealed recurrent copy number alterations in PIN and PCa lesions, and evidence of clonal relationships between PIN and invasive PCa. These studies provide new insight into PIN biology and the relationship between PIN and invasive PCa. Citation Format: Siri H. Strand, Okyaz Eminaga, Sujay Vennam, Chunfang Zhu, Jason Wang, Sushama Varma, Rosalie Nolley, Christian Kunder, Jonathan Pollack, Andreas Roeder, Karina D. Sorensen, James D. Brooks, Robert B. West. Molecular analysis of matched PIN, invasive prostate cancer, and adjacent normal prostate tissue samples reveal distinct transcriptional signatures and clonal relationships [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr A017.

Abstract A044: Generation of a new mouse model to study the role of oncofetal Cripto in aggressive lethal prostate cancer

Abstract Introduction: Early stages of prostate cancer (PCa) are commonly treated with surgery and androgen-deprivation therapy, but PCa can turn castration resistant due to pre-existing stem cell-like cells which might re-initiate tumor growth and lead to metastasis. Previously, it has been shown that human tumors express high levels of Cripto, an oncofetal protein, for this reason our hypothesis is that Cripto might play a role in PCa initiation and progression. We aim to study its oncogenic features in GEMMs models that are representative of early and late metastatic PCa. Methods: We performed conditional Cripto (CRIPTOflox/flox) knock out on N (Nkx3.1CreERT2, R26 LSL-YFP/LSL-YFP), NP (Nkx3.1CreERT2; Ptenflox/flox, R26 LSL-YFP/LSL-YFP) and NPK (Nkx3.1CreERT2; Ptenflox/flox; KrasLSL-G12D/+, R26 LSL-YFP/LSL-YFP) to generate respectively NC, NPC and NPKC. N animals mirror normal epithelium, whereas first stage of the disease, high-grade prostatic intraepithelial neoplasia (PIN)/carcinoma lesions with local invasive epithelium, are seen in NP. More advanced stage with invasive prostate adenocarcinoma with metastasis are developed in NPK. In vivo experiments’ workflow was the following: 8-weeks old mice were castrated and induced one month later with 5 daily injections of tamoxifen, after 2.5 weeks, animals were weekly treated with testosterone (10 weeks). Organoids were generated from YFP+/− single cell population recovered by FACS sorting. Results: Histopathological evaluation of newly generated NPC and NPKC showed presence of mPIN (100%, nNPC= 6, nNPKC=2), with a dominant cribriform morphology. NPKC additionally, features invasive PCa with an extent dominant pattern of 100%, showing portions with dense stroma forming whorl-like structures and occasional sheet-like accumulations of polygonal in stroma of regions with mPIN. In general, NPC and NPKC feature a more reactive stroma with a mild/moderate inflammation compared to the published models. OrganoidsYFP+ recapitulate molecular prostate tissue features, expressing luminal and basal markers. Organoids’ morphology varies consistently: N and NC are low in density and present a more cystic morphology, which is consistent with low-grade PIN phenotypes, whereas NP and NPK organoids are solid and denser which mimics an oncogenic transformation. In general, NC, NPC and NPKC organoids present a higher percentage of hollow organoids compared to N, NP and NPK respectively. Conclusions: NPC and NPKC phenotype specifically, showed important stromal alterations suggesting that Cripto might play a role not only on the epithelial compartment as well as in the stroma. For this reason, studies on secreted Cripto inhibition with ALK4-Fc are on-going. Our results show that organoids are an efficient in vitro model replicating different phenotypes seen in vivo. Citation Format: Elisa Rodrigues Sousa, Eugenio Zoni, Mario Scarpa, Marta De Menna, Allen Abey Alexander, Simone De Brot, George N. Thalmann, Marianna Kruithof-de Julio. Generation of a new mouse model to study the role of oncofetal Cripto in aggressive lethal prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr A044.

Identification of the Optimal Candidates for Nodal Staging with Extended Pelvic Lymph Node Dissection Among Prostate Cancer Patients Who Underwent Preoperative Prostate-specific Membrane Antigen Positron Emission Tomography. External Validation of the Memorial Sloan Kettering Cancer Center and Briganti Nomograms and Development of a Novel Tool

BackgroundAlthough the therapeutic role of extended pelvic lymph node dissection (ePLND) in patients with prostate cancer (PCa) is still under debate, this procedure is recommended for staging purposes in selected cases. Nomograms for predicting lymph node invasion (LNI) do not account for prostate-specific membrane antigen (PSMA) positron emission tomography (PET) imaging, which is characterized by a high negative predictive value for nodal metastases. ObjectiveTo externally validate models predicting LNI in patients with miN0M0 PCa at PSMA PET and to develop a novel tool in this setting. Design, setting, and participantsOverall, 458 patients with miN0M0 disease undergoing radical prostatectomy (RP) and ePLND at 12 centers between 2017 and 2022 were identified. Outcome measurements and statistical analysesAvailable tools were externally validated using calibration plots, the area under the receiver operating characteristic curve (AUC), and decision curve analyses to assess calibration, discrimination, and the net benefit. A novel coefficient-based model was developed, internally validated, and compared with available tools. Results and limitationsOverall, 53 patients (12%) had LNI. The AUC was 69% for the Briganti 2012, 64% for the Briganti 2017, 73% for the Briganti 2019, and 66% for the Memorial Sloan Kettering Cancer Center nomogram. Multiparametric magnetic resonance imaging stage, biopsy grade group 5, the diameter of the index lesion, and the percentage of positive cores at systematic biopsy were independent predictors of LNI (all p ≤ 0.04). Internal cross-validation confirmed a coefficient-based model with AUC of 78%, better calibration, and a higher net benefit in comparison to the other nomograms assessed. Use of a 5% cutoff would have spared 47% ePLND procedures (vs 13% for the Briganti 2019 nomogram) at the cost of missing only 2.1% LNI cases . The lack of central review of imaging and pathology represents the main limitation. ConclusionsTools for predicting LNI are associated with suboptimal performance for men with miN0M0 PCa. We propose a novel model for predicting LNI that outperforms available tools in this population. Patient summaryTools currently used to predict lymph node invasion (LNI) in prostate cancer are not optimal for men with negative node findings on PET (positron emission tomography) scans, leading to a high number of unnecessary extended pelvic lymph node dissection (ePLND) procedures. A novel tool should be used in clinical practice to identify candidates for ePLND to reduce the risk of unnecessary procedures without missing LNI cases.

Nigericin attenuates lipopolysaccharides induced sepsis and alleviates acute lung injury in in vivo model

Nigericin is a potent antibiotic derived from Streptomyces hygroscopicus which acts as a polyether ionophore. The anticancer property of nigericin was extensively studied, and it has been proven to inhibit cancer cell proliferation, progression, and invasion in prostate, colorectal, nasopharyngeal, and lung cancer. It also stimulates bacterial clearance in macrophages. The protective role of nigericin in sepsis, which is the foremost risk factor for acute lung injury and acute respiratory distress syndrome. Since bacteremia is frequently associated with lung injury, in this study we evaluated the potency of nigericin against an lipopolysaccharide (LPS)-induced sepsis mice model. BALB/c mice were challenged with intratracheal administration of LPS and simultaneously treated with 10 mg/kg and 20 mg/kg nigericin for 3 consecutive days. The mice were euthanized, and the lung tissue was excised to determine the lung wet/dry ratio. Bronchoalveolar lavage fluid was collected and assessed for protein content, LDH, and immune cell count. The antioxidant activity of nigericin was evaluated by measuring the MPO, MDA, SOD, and GSH activities in the lung tissues. The proinflammatory cytokines TNF-α and IL-6 were quantified to assess the anti-inflammatory potency against LPS-induced inflammation. iNOS, COX-2 and PGE-2 levels in lung tissue of nigericin-treated LPS-challenged mice were estimated to assess the immunomodulatory activity of nigericin. Finally, the antiapoptotic potency and cytoprotective effect of nigericin were analyzed by estimating apoptotic protein levels in lung tissue and assessing lung tissue histopathology with hematoxylin & eosin staining. Nigericin treatment significantly reduced immune cell infiltration, synthesis of proinflammatory cytokines, immunomodulatory proteins, and proapoptotic proteins in LPS-challenged mice. It also increased antioxidants and antiapoptotic protein Bcl2, thereby ameliorating lung tissue from LPS-induced inflammation. Our results indicate nigericin suppressed the LPS induced sepsis in an in vivo model and has the therapeutic potency to cure sepsis induced acute lung injury.

Development and internal validation of a novel nomogram for predicting lymph node invasion for prostate cancer patients undergoing extended pelvic lymph node dissection.

Few studies have focused on the performance of Briganti 2012, Briganti 2017 and MSKCC nomograms in the Chinese population in assessing the risk of lymph node invasion(LNI) in prostate cancer(PCa) patients and identifying patients suitable for extended pelvic lymph node dissection(ePLND). We aimed to develop and validate a novel nomogram based on Chinese PCa patients treated with radical prostatectomy(RP) and ePLND for predicting LNI. We retrospectively retrieved clinical data of 631 patients with localized PCa receiving RP and ePLND at a Chinese single tertiary referral center. All patients had detailed biopsy information from experienced uropathologist. Multivariate logistic-regression analyses were performed to identify independent factors associated with LNI. The discrimination accuracy and net-benefit of models were quantified using the area under curve(AUC) and Decision curve analysis(DCA).The nonparametric bootstrapping were used to internal validation. A total of 194(30.7%) patients had LNI. The median number of removed lymph nodes was 13(range, 11-18). In univariable analysis, preoperative prostate-specific antigen(PSA), clinical stage, biopsy Gleason grade group, maximum percentage of single core involvement with highest-grade PCa, percentage of positive cores, percentage of positive cores with highest-grade PCa and percentage of cores with clinically significant cancer on systematic biopsy differed significantly. The multivariable model that included preoperative PSA, clinical stage, biopsy Gleason grade group, maximum percentage of single core involvement with highest-grade PCa and percentage of cores with clinically significant cancer on systematic biopsy represented the basis for the novel nomogram. Based on a 12% cutoff, our results showed that 189(30%) patients could have avoided ePLND while only 9(4.8%) had LNI missing ePLND. Our proposed model achieved the highest AUC (proposed model vs Briganti 2012 vs Briganti 2017 vs MSKCC model: 0.83 vs 0.8 vs 0.8 vs 0.8, respectively) and highest net-benefit via DCA in the Chinese cohort compared with previous nomograms. In internal validation of proposed nomogram, all variables had a percent inclusion greater than 50%. We developed and validated a nomogram predicting the risk of LNI based on Chinese PCa patients, which demonstrated superior performance compared with previous nomograms.

Bacterial lipopolysaccharide-related genes are involved in the invasion and recurrence of prostate cancer and are related to immune escape based on bioinformatics analysis.

The composition of the tumor microbial microenvironment participates in the whole process of tumor disease. However, due to the limitations of the current technical level, the depth and breadth of the impact of microorganisms on tumors have not been fully recognized, especially in prostate cancer (PCa). Therefore, the purpose of this study is to explore the role and mechanism of the prostate microbiome in PCa based on bacterial lipopolysaccharide (LPS)-related genes by means of bioinformatics. The Comparative Toxicogenomics Database (CTD) was used to find bacterial LPS- related genes. PCa expression profile data and clinical data were acquired from TCGA, GTEx, and GEO. The differentially expressed LPS-related hub genes (LRHG) were obtained by Venn diagram, and gene set enrichment analysis (GSEA) was used to investigate the putative molecular mechanism of LRHG. The immune infiltration score of malignancies was investigated using single-sample gene set enrichment analysis (ssGSEA). Using univariate and multivariate Cox regression analysis, a prognostic risk score model and nomogram were developed. 6 LRHG were screened. LRHG were involved in functional phenotypes such as tumor invasion, fat metabolism, sex hormone response, DNA repair, apoptosis, and immunoregulation. And it can regulate the immune microenvironment in the tumor by influencing the antigen presentation of immune cells in the tumor. And a prognostic risk score and the nomogram, which were based on LRHG, showed that the low-risk score has a protective effect on patients. Microorganisms in the PCa microenvironment may use complex mechanism and networks to regulate the occurrence and development of PCa. Bacterial lipopolysaccharide-related genes can help build a reliable prognostic model and predict progression-free survival in patients with prostate cancer.

Circular RNA circARHGEF28 inhibited the progression of prostate cancer via the miR-671-5p/LGALS3BP/NF-κB axis.

Circular RNAs (circRNAs) play crucial roles in various biological processes, including prostate cancer (PCa). However, the precise roles and mechanism of circRNAs are complicated. Hence, we studied the function of a circRNA that might be involved in the progression of PCa. In this study, we found that circARHGEF28 was frequently downregulated in PCa tissues and cell lines. Furthermore, gain- and loss-of function experiments in vitro showed that circARHGEF28 inhibited proliferation, migration, and invasion of PCa. Additionally, circARHGEF28 suppressed PCa progression in vivo. Bioinformatics analysis and RNA pull-down and capture assay found that circARHGEF28 sponged miR-671-5p in PCa cells. Importantly, qRT-PCR and dual luciferase assays found that Lectin galactoside-binding soluble 3 binding protein (LGALS3BP) was downstream of miR-671-5p, and western blot analysis further confirmed that LGALS3BP negatively regulated the nuclear factor kappa-B (NF-κB) pathway. These results demonstrated that circARHGEF28 abolished the degradation of LGALS3BP by sponging miR-671-5p, thus blocking the activation of the NF-κB pathway. Our findings revealed that circARHGEF28/miR-671-5p/LGALS3BP/NF-κB may be an important axis that regulates PCa progression.

PIM1 phosphorylates ABI2 to enhance actin dynamics and promote tumor invasion.

Distinguishing key factors that drive the switch from indolent to invasive disease will make a significant impact on guiding the treatment of prostate cancer (PCa) patients. Here, we identify a novel signaling pathway linking hypoxia and PIM1 kinase to the actin cytoskeleton and cell motility. An unbiased proteomic screen identified Abl-interactor 2 (ABI2), an integral member of the wave regulatory complex (WRC), as a PIM1 substrate. Phosphorylation of ABI2 at Ser183 by PIM1 increased ABI2 protein levels and enhanced WRC formation, resulting in increased protrusive activity and cell motility. Cell protrusion induced by hypoxia and/or PIM1 was dependent on ABI2. In vivo smooth muscle invasion assays showed that overexpression of PIM1 significantly increased the depth of tumor cell invasion, and treatment with PIM inhibitors significantly reduced intramuscular PCa invasion. This research uncovers a HIF-1-independent signaling axis that is critical for hypoxia-induced invasion and establishes a novel role for PIM1 as a key regulator of the actin cytoskeleton.

Abstract 1326: Perineural invasion and bone metastases in prostate cancer - is there a link

Abstract Bone metastases are the most common secondary lesions in prostate cancer (PCa) patients. Typically, initial bone metastases develop in the lower part of the spine and pelvis. While this metastatic pattern is not consistent with the random nature of the arterial osseous dissemination, previous reports suggested the role of the retrograde venous spread in this phenomenon. However, clinical data also indicate a positive association between the degree of perineural invasion (PNI) and bone metastases in PCa patients. Yet, the mechanism linking these two means of dissemination remains unknown. To identify the processes involved in the initial PCa dissemination we used the PC3-ML cell line, which was previously shown to trigger bone metastases, in the orthotopic xenograft model. To this end, the cells were injected into the prostates of male SCID/Bg mice and tumor development and progression was followed by magnetic resonance imaging (MRI) and histopathology. MRI indicated local invasion of the primary tumor, which later resulted in the formation of multiple secondary lesions along the lower spine. However, histopathological analysis revealed that these lesions were initiated in the soft tissues surrounding the vertebral column, while the subsequent invasion of the vertebra was the secondary event. The staining for neuronal markers, neurofilament (NF), tyrosine hydroxylase (TH) and neuropeptide Y (NPY), revealed that PCa cells created multicellular clusters spreading longitudinally along the small nerve fibers, particularly in the paraspinal muscle compartment. The staining for the endothelial cell marker, CD31, detected small vessels present within PCa clusters. However, no evident encapsulation of the PCa cells within blood vessels consistent with the retrograde venous spread was observed. Altogether, these results suggest PNI as the main route of the initial local dissemination of the PCa cells toward the bone, which then triggers bone invasion and subsequent hematogenous spread to distant bones. Further mechanistic studies are required to confirm this hypothesis. Citation Format: Dawid Sigorski, Susana Galli, Sung Hyeok Hong, Ewa Iżycka-Świeszewska, Joanna B. Kitlinska. Perineural invasion and bone metastases in prostate cancer - is there a link [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 1326.

Abstract 2564: Targeting IGF-1/STAT3 signaling crosstalk to inhibit prostate cancer growth and metastasis

Abstract Introduction: Prostate cancer is the most prevalent cancer in American men with unknown etiology. Signal transducer and activator of transcription 3 (STAT3) is a transcription factor, which has been implicated in the growth and metastasis of prostate cancer. Activation of STAT3 through phosphorylation modulates the expression of miR-21, an oncomir, by directly binding to their promoter region in response to various cytokines and growth factors. One such growth factor is insulin-like growth factor (IGF)-1, whose elevated levels are linked to the growth of prostate cancer. IGF-1 exerts its mitogenic effect by binding to its receptor, IGF-1R, whose expression on prostate cancer cells is also increased during neoplastic transformation. There is limited information concerning the crosstalk between IGF-1/IGF-1R and STAT3 signaling pathways which could explain their oncogenic roles in prostate cancer. In this study, we examined whether STAT3-mediated increase of miR-21 is critical for the proliferative effects of IGF-1. We further show that resveratrol, a polyphenolic antioxidant, attenuates prostate cancer growth and invasiveness through regulation of IGF-1/STAT3 pathway. Methods: In vitro studies were performed on DU145 cells, which are androgen receptor negative human prostate cancer cells. Cell viability was assessed by MTS assays, while invasion and migration of cells were tested by Boyden chamber assay and wound healing assay, respectively. Modulation of protein expression was determined by Western blotting. MiR-21 levels were determined by TaqMan assay. SiRNA transfections were performed to suppress the expression of specific proteins in order to elucidate the molecular mechanisms underlying the actions of IGF-1, STAT3 and resveratrol. Results: Acute IGF-1 treatment of DU145 cells increased phosphorylation of Akt which was correlated with the activation of STAT3. This effect of IGF-1 was through activation of IGF-1R, as siRNA-mediated knockdown of IGF-1R attenuated STAT3 activation and Akt phosphorylation. IGF-1 treatment also increased miR-21 levels and decreased the expression of its downstream protein, programmed cell death 4 (PDCD4). Effects of IGF-1 were completely attenuated by resveratrol. Furthermore, knockdown of IGF-1R and STAT3 mimicked the actions of resveratrol on DU145 cells suggesting that a signaling crosstalk exists between IGF-1 and STAT3 pathways which can be targeted to inhibit prostate cancer. Conclusion: This study demonstrates, for the first time, that IGF-1 treatment significantly activates STAT3 pathway and increases miR-21 expression in prostate cancer cells. Targeted inhibition of IGF-1-induced activation of STAT3 by resveratrol holds the promise for the development of novel therapeutic agents against prostate cancer. Citation Format: Sandeep Sheth, Raheem Al-Aameri, Vickram Ramkumar. Targeting IGF-1/STAT3 signaling crosstalk to inhibit prostate cancer growth and metastasis [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 2564.

Abstract 3631: Infiltrating Bruton’s tyrosine kinase expressed B cells effects on prostate cancer metastasis

Abstract Prostate cancer is the most common type of cancer found in senior males and is the 2nd leading cause of cancer death in American males. Although prostate cancer grows slowly within the prostate glands, because of a lack of efficient treatments at the appropriate timing, cancer cells eventually become aggressive and metastatic. The progression of cancer is supported by many factors, such as signals received from surrounding cells, like B cells and other effector cells. B cells play a role as a part of the adaptive immune system to produce antibodies. To become mature B cells, they rely on activation of a non-receptor kinase, Bruton’s Tyrosine Kinase (BTK). Preliminary data from our lab showed that in the prostate cancer tissues, more infiltrated B cells expressing BTK were present nearby the prostate cancer cells. Upon this finding, we hypothesize that BTK expressed B cells will support the survival of prostate cancer cells through BTK signaling. In this study, we will overexpress BTK in B cells to study the proliferation and migration of prostate cancer cells through a co-culture system. Our findings will show an increase in migration of prostate cancer cells when cultured with BTK expressed B cells. In the future, we will validate the importance of BTK in regulating prostate cancer migration and invasion; and identify their mechanisms. Our overall findings from this project would reveal key elements of the signaling pathways of BTK expressed B cells to promote prostate cancer migration. Citation Format: Crystal Byrd, Geou- Yarh Liou. Infiltrating Bruton’s tyrosine kinase expressed B cells effects on prostate cancer metastasis. [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 3631.

Grading Intraductal Carcinoma of the Prostate; ISUP versus GUPS Recommendations.

To evaluate the frequency of intraductal component (IDC-P) in prostatic adenocarcinoma and its effect on the final grade using the ISUP and GUPS grading system. Descriptive study. Place and Duration of the Study:Shaukat Khanum Memorial Cancer Hospital and Research Centre, Lahore, between June 2021 and June 2022. The study included 250 cases of prostatic adenocarcinoma. The presence of the intraductal carcinoma prostate (IDC-P) was confirmed by patchy or complete staining of the basal cell layer by p63 immunohistochemical stain. Cases with IDC-P were then graded using two different methods, first using the grading criteria based on the ISUP recommendations and then by the grading criteria based on the GUPS recommendations. Two hundred and fifty cases showed invasive prostatic carcinoma ranging from Gleason grade group 2-5. IDC-P was identified in 5 of the 250 biopsies (2%). The final Gleason grade remained unchanged in these cases, when they were graded using the ISUP and GUPS recommendations. Although the present results are based on a relatively small sample size, IDC-P was not frequently present in biopsies of patients with adenocarcinoma in the studied population. Grading IDC-P in invasive prostate cancer led to only a minor change in grade group assignment of prostate cancer biopsies. Prostatic adenocarcinoma, Intraductal carcinoma, IDC-P, ISUP, GUPS, Gleason Grade group.

MP20-14 GENERATION OF A NEW GEMMs MODEL TO EXPLORE THE ROLE OF ONCOFETAL CRIPTO IN PROSTATE CANCER PROGRESSION

You have accessJournal of UrologyCME1 Apr 2023MP20-14 GENERATION OF A NEW GEMMs MODEL TO EXPLORE THE ROLE OF ONCOFETAL CRIPTO IN PROSTATE CANCER PROGRESSION Elisa Rodrigues Sousa, Eugenio Zoni, Mario Scarpa, Marta De Menna, Allen Abey Alexander, Simone De Brot, George N. Thalmann, and Marianna Kruithof-de Julio Elisa Rodrigues SousaElisa Rodrigues Sousa More articles by this author , Eugenio ZoniEugenio Zoni More articles by this author , Mario ScarpaMario Scarpa More articles by this author , Marta De MennaMarta De Menna More articles by this author , Allen Abey AlexanderAllen Abey Alexander More articles by this author , Simone De BrotSimone De Brot More articles by this author , George N. ThalmannGeorge N. Thalmann More articles by this author , and Marianna Kruithof-de JulioMarianna Kruithof-de Julio More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003245.14AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Early stages of prostate cancer (PCa) are treated with surgery and androgen-deprivation therapy, but PCa can turn castration resistant possibly due to stem cell-like cells that re-initiate PCa growth and lead to metastasis. Given that Cripto is highly expressed in human tumors, we hypothesise that Cripto might play a role in tumor initiation and late progression, so we knockout Cripto in PCa GEMMs models with the aim of studying its oncogenic role. METHODS: N ( Nkx3.1CreERT2, R26 LSL-YFP/ LSL-YFP), NP (Nkx3.1CreERT2; Ptenflox/flox, R26 LSL-YFP/ LSL-YFP) and NPK (Nkx3.1CreERT2; Ptenflox/flox; KrasLSL-G12D/+, R26 LSL-YFP/ LSL-YFP) were crossed with conditional Cripto knockout (CRIPTOflox/flox) to generate NC, NPC and NPKC. N mice mirror normal epithelium, NP mimic the first stage of the disease, with high-grade prostatic intraepithelial neoplasia (PIN)/carcinoma lesions with local invasive epithelium, while NPK have invasive prostate adenocarcinoma with metastasis. In vivo experiments’ workflow is the following: castration of 8-weeks old mice and induction after 1 month with 5 daily injections of tamoxifen, after 2.5 weeks mice were treated with weekly testosterone (10 weeks). Single cells were isolated from prostate tissue and the YFP+/- population recovered by FACS sorting and cultured as organoids. RESULTS: Histopathological evaluation of NPC and NPKC showed presence of mPIN (100%) with a dominant cribriform morphology. NPKC feature invasive PCa with an extent dominant pattern of 100% and portions with dense stroma forming whorl-like structures and occasional sheet-like accumulations of polygonal in mPIN regions. NPC and NPKC, compared to NP and NPK, present a more reactive stroma with a mild/moderate inflammation reaction. Organoids express luminal and basal markers mirroring molecular features of matched tissues. Morphology varies consistently: NP, NPC, NPK and NPKC organoids are solid which is consistent with an oncogenic transformation, whereas N and NC organoids present a cystic morphology, with lower densities, which correlates with low-grade PIN phenotypes. NC, NPC and NPKC organoids feature a higher percentage of hollow organoids compared to N, NP and NPK respectively. CONCLUSIONS: Stromal alterations in NPC and NPKC might suggest a role for Cripto not only restricted to prostate epithelium. For this reason, studies on secreted Cripto inhibition with ALK4-Fc are planned. Our finding support that organoids are an efficient in vitro model replicating in vivo phenotypes. Source of Funding: United States Department of Defense © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e281 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Elisa Rodrigues Sousa More articles by this author Eugenio Zoni More articles by this author Mario Scarpa More articles by this author Marta De Menna More articles by this author Allen Abey Alexander More articles by this author Simone De Brot More articles by this author George N. Thalmann More articles by this author Marianna Kruithof-de Julio More articles by this author Expand All Advertisement PDF downloadLoading ...

Sympathetic β2-adrenergic receptor blockade overcomes docetaxel resistance in prostate cancer

PurposeDue to the limited effective therapies, resistance to docetaxel is ordinarily fatal and remains a critical clinical challenge.β2-adrenergic receptor(β2-AR)can promote the metastasis and invasion of prostate cancer, but the role in chemotherapy-resistant prostate cancer remains unclear. MethodsBy downloading the GEO database in NCBI, the expression of β2-AR in different prostate tissues was analyzed. We constructed docetaxel-resistant prostate cancer cell lines by the method of dose-escalation. LC3B-labeled stable cells and shAtg5 knockdown stable cells were constructed by lentivirus infection. The molecular mechanism of β2-AR affecting docetaxel sensitivity through apoptosis and autophage were investigated by flow cytometry, mitochondrial membrane potential and western blot. Then we detected the interaction between autophagy and apoptotic by performing immunoprecipitation assay. ResultsWe show that restraining the activity of β2-AR sensitized the cell response and reduced the resistance to docetaxel. The mechanism involves the regulation of β2-AR in the cellular response to docetaxel through apoptosis and autophagy via caspase signaling and Atg5/AMPK/mTOR pathway as well as the effect of β2-AR on the crosstalk between apoptosis and autophagy via p38 MAPK and JNK/c-Jun/FOXO3a signaling pathways. ConclusionOur data demonstrate that β2-AR inhibitor-induced autophagy and apoptosis contribute to the effectiveness responses to docetaxel in castration-resistant prostate cancer, and in combination with pharmacological agents of β2-AR and autophagy inhibitors may provide a potential therapeutic strategy to enhance the limited capacity of docetaxel to control castration-resistant prostate cancer.