Assessment of BRD4 intermolecular interaction

Approximately 50% of prostate cancer (PCa) patients harbor fusions involving the TMPRSS2 and ERG genes. Despite this, tailored therapies targeting the fused gene, tERG, remain undeveloped. Our study analyzed biopsy samples from two clinical trials assessing the efficacy of androgen receptor (AR) signaling inhibitors (ARSIs). The results revealed that tERG promotes resistance to ARSIs and is associated with elevated levels of the glucocorticoid receptor (GR). Subsequent assays showed that GR directly interacts with tERG, alleviates allosteric autoinhibition, and prevents chemotherapy-induced tERG degradation. In PCa models, either inhibiting GR or lowering cortisol levels suppressed tumor growth in tERG-positive models, but not in tERG-negative models. In addition, patient-derived fusion-positive xenografts displayed enhanced sensitivity to combined GR and AR inhibitors. Collectively, these findings highlight TMPRSS2-ERG as a new biomarker and propose that simultaneous inhibition of GR and AR may specifically benefit tERG-positive patients. However, GR stimulatory corticosteroid therapies may not be advisable for this patient subgroup.

Computational prediction of BRD4 intermolecular interaction and co-IP validation

Sequences for primers used for qPCR analysis

BRD4S depletion attenuates migrative and invasive ability of CDK12-deficient cancer cells

Immunotherapy has been effective in many cancer types but has failed in multiple clinical trials in prostate cancers, with the underlying mechanisms remaining largely unclear. Here, we demonstrate that androgen receptor pathway inhibitor (ARPI) plus irradiation (IR) triggered robust anticancer immunity in prostate cancers in both patients and mice. We show that androgen-activated AR suppressed innate immune signaling by inducing inhibitor of nuclear factor kappa-B kinase subunit epsilon (IKBKE) gene repression through HDAC2 interaction with an IKBKE enhancer RNA (IKBKE eRNA, or IKBKE-e). ARPI treatment caused IKBKE derepression and enhanced an IR-induced innate immune response via action of RIG-I and MDA5 dsRNA sensors. IKBKE-e ablation largely enhanced innate immunity in prostate cancer cells in culture and anticancer immunity in mice. Our results revealed AR, HDAC2, and IKBKE eRNA as critical intrinsic immune suppressors in prostate cancer cells, suggesting that rejuvenating inhibitor of nuclear factor kappa-B kinase subunit epsilon (IKKε) signaling by targeting IKBKE-e is an actionable strategy to elicit synthetic anticancer immunity in immunologically "cold" cancers such as prostate cancer.

In diagnosing type 1 autoimmune pancreatitis (AIP), serum IgG4 (sIgG4) can be false-negative. EUS-guided fine-needle aspiration/biopsy (EUS-FNA/FNB) pathology is key for diagnosis, but clinical features’ impact on pathologic confirmation is unclear. This study analyzed their link and factors improve diagnostic accuracy. We analyzed data from a single-center retrospective study at Changhai Hospital (Jan 2009-Jan 2024). Type 1 AIP was diagnosed per International Consensus Diagnostic Criteria (ICDC). Patients with surgical diagnosis, no EUS, or incomplete biopsy data were excluded; eligible cases were grouped into “Confirmed”/“Unconfirmed” per ICDC. Baseline data, laboratory indicators, imaging, and EUS-FNA/FNB data were collected. Statistical analyses (ROC, χ² tests, multivariate logistic regression) were done with R 4.4.0. A total of 182 suspected type 1 AIP patients were enrolled; 84.07

CDK12 inhibition, but not CDK13, increases BRD4S mRNA expression

BRD4S promotes cancer migration and invasion in vitro

Elucidating the structure and function of enteropeptidase (EP) is essential for advancing our understanding of its biological significance, particularly in regulating trypsinogen activation. Using cryo-EM and enzymatic activity, we uncovered the significance of the CUB2 domain in mediating the cleavage of macromolecular substrates. We identified crucial binding loops and key residues for EP's proteolytic function. The mutation E574A enhanced the proteolytic activity of EP, whereas the mutation N619A diminished its cleavage efficiency, highlighting the importance of surface-charged interactions in modulating EP's activity. A proteolytic cycle was proposed to deepen our understanding of the trypsinogen activation by EP. This work offers valuable insights into the molecular mechanisms underlying EP's interaction with its substrate, and opens up avenues for therapeutically modulating EP-mediated proteolysis.

In the research on the pathogenesis of post-acute pancreatitis diabetes mellitus (PPDM-A), abnormal bidirectional communication between pancreatic acinar cells (PACs) and islet cells is a key focus. However, immortalized cell lines cannot replicate pathophysiological conditions, making the extraction of high-quality primary cells crucial. Current methods for extracting primary islets from mice mostly rely on in-vivo pancreatic perfusion via bile duct cannulation, which has a high technical barrier and is not conducive to operation by researchers without experience. This study modified the method, eliminating the need for complex, in-vivo perfusion. SPF-grade C57BL/6J mice (6-8-week-old) were anesthetized and euthanized, followed by pancreas isolation. The pancreas was digested in vitro with collagenase P; primary islets were separated via Ficoll density gradient centrifugation, and acinar cells were obtained through cell sieve filtration and centrifugation. Cell viability and function were evaluated using calcein/propidium iodide (Calcein/PI) staining, glucose-stimulated insulin secretion assay, and amylase activity detection. The results showed that the modified method was easy to operate: the yield per mouse was (120 ± 5) primary islets and 1.6-1.95 × 10⁷ acinar cells; the viability rates of islets and acinar cells were (97.52 ± 0.16)% and (96.55 ± 0.95)%, respectively. Moreover, the islets exhibited normal insulin secretion ability, and the acinar cells were sensitive to cerulein stimulation. This method is simple and reliable, providing a feasible framework for studying pancreatic exocrine-endocrine interactions and PPDM-A. However, it has limitations, such as an unvalidated application in rats.

BRD4 inhibitors inhibit CDK12 deficiency induced cancer cell migration and invasion

Aberrant tumor metabolism plays a crucial role in establishing and sustaining an immunosuppressive microenvironment. CD73, a key enzyme in the purine metabolism pathway, is frequently overexpressed in tumors, leading to elevated extracellular adenosine (ADO) levels. This accumulation suppresses antitumor immune responses by promoting immune cell exhaustion, ultimately contributing to immunotherapy resistance. However, the mechanisms underlying CD73 upregulation in tumors remain poorly defined. Here, we identify aurora kinase B (AURKB) as a critical regulator of CD73 overexpression in renal cell carcinoma (RCC), orchestrating immune evasion and resistance to immune checkpoint therapy. AURKB is highly expressed in RCC, with its prognostic significance being especially evident in tumors with strong immunogenicity. Mechanistically, AURKB enhances CD73 expression, thereby promoting T cell dysfunction and immune suppression. Mass cytometry (CyTOF) analysis revealed that AURKB inhibition facilitates immune infiltration and alleviates immune exhaustion. We further demonstrate that AURKB directly interacts with CD73 and phosphorylates it at serine 429, which inhibits STUB1-mediated ubiquitination and proteasomal degradation of CD73. Targeting AURKB to modulate the post-translational stability of CD73 restores antitumor immunity and synergizes with immune checkpoint blockade in vivo. Collectively, this study uncovers a previously unrecognized AURKB-CD73 axis and provides a mechanistically informed strategy to enhance immunotherapy efficacy in RCC.

Chronic pancreatitis is a progressive inflammatory disease, whose hallmark pathological feature is an irreversible fibrosis process. This process further leads to irreversible structural damage, pancreatic functional decline, and a variety of severe complications. Common complications mainly include pancreatic exocrine insufficiency, endocrine insufficiency, metabolic bone disease, pain, pancreatic duct stones, obstruction of common bile duct and duodenum, pancreatic pseudocysts, and vascular complications. This review systematically sorts out and integrates the etiology, diagnostic criteria, and management of various complications. Moreover, active intervention for these complications through secondary prevention and standardized management is crucial for improving patients’ clinical prognosis. Ultimately, this review establishes a comprehensive “Etiology-Diagnosis- Management “clinical reference system for the primary complications of chronic pancreatitis, providing gastroenterologists with a quick reference for the standardized diagnosis and management of chronic pancreatitis-related complications.