Renal cell carcinoma (RCC) is a heterogeneous malignancy for which clear cell RCC (ccRCC) represents the most common and clinically aggressive subtype. Survivin (encoded by BIRC5), an inhibitor of apoptosis and key regulator of mitosis, is frequently overexpressed in RCC and associated with poor prognosis, yet its broader role in kidney cancer biology remains poorly defined. By analyzing transcriptomic data from The Cancer Genome Atlas-kidney renal clear cell carcinoma (TCGA-KIRC) cohort, we found that advanced-stage ccRCC exhibits widespread dysregulation of cell-cycle pathways, with 15,160 genes upregulated and 479 genes downregulated in stage IV compared to stage I tumors. To define survivin's functional contribution, we performed loss-of-function and pharmacologic inhibition studies in murine RENCA and human 786-O RCC cell lines. Survivin knockdown or treatment with the small-molecule inhibitor YM155 reduced proliferation, S-phase entry and cyclin D1 expression, and impaired collective (wound-healing), single-cell and transwell migration. Unexpectedly, survivin depletion increased mitochondrial content while lowering oxygen consumption, indicating accumulation of dysfunctional mitochondria. Together, these findings identify survivin as a node between cell-cycle progression and mitochondria in RCC.

Glioblastoma Multiforme (GBM) is a highly aggressive brain cancer characterized by rapid proliferation and extensive remodeling of the extracellular matrix (ECM), leading to progressive tissue stiffening. Although ECM stiffness is known to promote GBM progression, the molecular mechanisms linking mechanical cues to tumor growth remain insufficiently defined. In this study, transcriptomic comparison of GBM tumors and non-neoplastic brain tissue revealed coordinated upregulation of cell cycle regulators and matrisome-associated genes, with survivin (BIRC5) identified as a central node linking proliferative signaling and ECM remodeling networks. Analysis of GBM patient specimens further showed strong nuclear survivin expression in regions with elevated collagen deposition. To directly evaluate stiffness-dependent regulation of survivin, GBM cells were cultured on fibronectin-infused hydrogels with tunable stiffness. Stiff matrices increased survivin expression along with cyclin D1 and cyclin A, consistent with increased cell cycle progression. Pharmacologic inhibition or siRNA-mediated suppression of survivin reduced stiffness-induced proliferation and attenuated expression of matrisome components, including collagens and lysyl oxidase. These findings indicate that survivin functions as a mechanosensitive regulator that coordinates cell cycle progression with ECM production in stiff tumor microenvironments. Collectively, this study identifies survivin as a key mediator linking ECM stiffness to GBM growth and matrisome remodeling. Targeting survivin and its effectors may offer a mechanosensitive strategy to limit GBM growth.

Pathologic arterial stiffening is a hallmark of vascular disease that contributes to maladaptive vascular remodeling and neointimal hyperplasia through vascular smooth muscle cell (VSMC) phenotypic switching. Yet, because vascular disease progression is governed by both biomechanical and extracellular matrix (ECM) alterations, existing in vitro models often fail to recapitulate the full complexity of the diseased vascular microenvironment. Here, we developed a bioactive decellularized extracellular matrix (dECM) and methacrylated hyaluronic acid (MeHA) composite scaffold platform with tunable stiffness that preserves native vascular ECM components while enabling controlled investigation of stiffness-dependent cell behavior. Proteomic analyses confirmed retention of key vascular matrisome components, including collagens and glycoproteins, following decellularization. Electrospun vascular dECM scaffolds maintained an aligned fibrous architecture and spanned stiffness ranges representative of healthy and pathologically stiffened arterial microenvironments. Within this matrix-preserving platform, human VSMCs cultured on stiff dECM scaffolds exhibited increased spreading, altered morphology, enhanced nuclear localization of YAP and survivin, and broad transcriptional changes consistent with a shift toward a proliferative, matrix-remodeling VSMC phenotype. Together, this bioactive, matrix-preserving platform enables mechanobiologically relevant modeling of stiffness-driven vascular remodeling and indicates YAP and survivin as candidate regulators of maladaptive VSMC mechanotransduction.

Proteostasis declines with lung aging, while the role of the Unfolded Protein Response (UPR) in lung aging and age-associated pulmonary diseases remains understudied. We investigated how deficiency in the UPR sensor ATF6α affects physiological and smoke exposure-accelerated lung aging. ATF6α -deficient mice exhibited accelerated alveolar simplification, a sign of lung parenchymal aging, which was exacerbated by smoking. Nevertheless, small airway vascular fibrotic remodeling, a prominent smoking induced pathology, was not evident in smoke-exposed ATF6α -deficient mice. Mechanistically, these divergent phenotypes arose from cell-type-specific ATF6α programs. In alveolar epithelial type 2 cells (AEC2s), the facultative progenitors of the lung parenchyma, ATF6α maintained mitochondrial bioenergetics and sustained efficient re-differentiation into alveolar epithelial type 1 cells (AEC1s). In lung pericytes, ATF6α promoted extravasation, re-differentiation into myofibroblast-like cells, and production of collagens 1 and 3. These findings identify ATF6α as a cell-type-specific regulator of differentiation programs during lung aging and highlight the need to study ATF6α under defined physiological and pathological contexts before therapeutically targeting this pathway.

The 3D genome architecture is a higher-order organization of chromosomes within the nucleus that is critical to the control of epigenomic modifications. However, our knowledge regarding the role of 3D genomic organization in the regulation of vascular gene expression and function is lacking. In the present study, CRISPR-engineered rats modelled after two common polymorphisms (S188F and N126D) in human glucose-6-phosphate dehydrogenase (G6PD) revealed modifications to the 3D genome in aortas from rats expressing a deficient G6PD variant (S188F), but not a non-deficient one (N126D), is associated with: 1] up-regulated expression of TET enzymes that augmented expression of genes encoding antiproliferative proteins, 2] suppressed expression of genes encoding inflammatory/thrombotic/fibrotic proteins, and 3] reduced angiotensin II-induced aortic stiffness and hypertension. G6PD interacted with MATRIN-3, a nuclear matrix/scaffold protein, and a deficient G6PD variant increased the relative abundance of MATR3 and CCCTC-binding factors, potentially modifying 3D-genome structure. Additionally, G6PD deficiency-induced enrichment of H3K27ac likely influences the establishment and maintenance of the 3D genome. Therefore, we propose that the nexus between metabolism and the 3D genome regulates arterial gene expression and vascular disease.

The organization of the cell's cytoskeletal filaments is coordinated through a complex network of signaling cascades activated by both internal and external cues. Two major actin regulatory pathways are signal transduction through Rho family GTPases and growth and proliferation signaling through the Hippo pathway. These two pathways define the actin cytoskeleton, controlling foundational cellular attributes such as morphology and the organization of actin-based structures, and are hijacked to promote proliferation and motility in aggressive cancers. In this study, we use human epithelial cells to investigate the interplay between the Hippo and Rho Family signaling pathways. We identify that the RhoA GTPase-activating protein, ARHGAP18, forms a complex with two Hippo pathway components, the tumor suppressor Merlin (NF2), and the transcriptional coactivator YAP. Using super-resolution STORM microscopy, we characterize single-filament-level changes in the actin cytoskeleton that arise from CRISPR/CAS9 knockout of ARHGAP18. We report that the loss of ARHGAP18 results in cytoskeletal alterations associated with dysregulation of RhoA signaling at apical structures and aberrant nuclear localization of YAP. These findings provide additional support for models suggesting that Hippo and Rho family GTPase signaling cascades may be temporally and spatially coordinated in the regulation of the actin cytoskeleton.

The rotator cuff plays a vital role in shoulder movement and joint stability. Unfortunately, tears at the rotator cuff enthesis are common and frequently lead to retears after surgical intervention, particularly at the suture location and its anchor sites. These failures are typically due to the inability of current surgical treatments to mimic the native tissue complexity and provide the necessary metabolic, bioactive, and biophysical cues for effective enthesis regeneration. In this study, we engineered a biomimetic multiphasic scaffold system (BMS) to integrate with conventional suture anchor systems and deliver spatially organized structural and biological cues to enhance enthesis regeneration. The BMS consists of three distinct phases: Phase 1 features an aligned, nanofibrous decellularized tendon extracellular matrix (dECM) combined with “stiff” methacrylated hyaluronic acid (MeHA); phase 2 incorporates nonaligned, nanofibrous dECM with “soft” MeHA; and phase 3 uses a porous, bioenergetic, citrate-based composite scaffold for bone integration. In vitro, the BMS notably enhanced tenogenic, fibrochondrogenic, and chondrogenic differentiation, facilitating zone-specific rotator cuff enthesis regeneration. Further, in vivo, the BMS promoted successful integrative healing, forming distinct tendon, fibrocartilage, and bone regions at the repair site. This advanced multiphasic scaffold closely replicates native tissue properties, offering a promising strategy to improve rotator cuff repair. Its integration with conventional suture anchors provides an innovative design that enhances mechanical fixation and guides enthesis healing to reduce retear rates. Broadly, this platform offers a versatile solution for biointegrative repair strategies across complex soft-to-hard tissue interfaces.

Introduction: Stroke remains the 5 th leading cause of death worldwide, yet its mechanisms and treatment options remain limited. Intracranial atherosclerotic disease, characterized by plaque formation in cerebral vessels, contributes to ischemic stroke, and both plaque stiffness and extracellular matrix (ECM) remodeling play critical roles in stroke pathology. Survivin ( BIRC5 ), a pro-proliferative&anti-apoptotic protein, is upregulated in pathological (stiff) vascular conditions like atherosclerosis, stroke, and hypertension, all of which are associated with increased stiffness. Survivin promotes plaque development, and is linked to poor cardiovascular outcomes. Although direct connections between survivin and stroke are limited, vascular stiffness remains a key risk factor. Methods: We analyzed a public RNA-seq dataset (GSE137482) of cerebral arteries from 18-month-old mice subjected to cerebral ischemia using the middle cerebral artery occlusion model ( Fig 1A ). Given that dysregulated vascular smooth muscle cells (VSMCs) are linked to vascular stiffening and survivin upregulation, both observed in ischemic stroke, we performed RNA-seq on human VSMCs with survivin knockdown cultured on a stiff hydrogel mimicking the mechanical properties of atherosclerosis or stroke-associated environments. Results: Analysis of the stroke dataset revealed enrichment of ECM-related terms in the Gene Ontology (GO) cellular component category, while biological processes were associated with signaling, development, and response to stimuli. Given that survivin modulates ECM synthesis and stroke-induced mice showed strong ECM enrichment, we analyzed matrisome gene expression and observed differential expression of collagens ( Fig 1B ), proteoglycans ( Fig 1C ), and ECM glycoproteins ( Fig 1D ) compared to controls. To examine survivin’s role, we compared the stroke dataset with the VSMC dataset ( Fig 2A ) and found that survivin inhibition dramatically altered both matrisome composition and proliferative pathways ( Fig 2B-D ), underscoring its therapeutic potential. Finally, Ingenuity Pathway Analysis (IPA) of the survivin knockdown dataset predicted mitigation of blood–brain barrier disruption and ischemic stroke, key processes in stroke pathology, through genes including APP, STAT3, and PLAT ( Fig 3A-B ). Conclusion: These findings suggest mechanistic links between survivin, vascular stiffness, and stroke, identifying potential targets for therapeutic strategies.

Abstract Clear cell renal cell carcinoma (ccRCC) is the most aggressive subtype of renal cell carcinoma. TCGA analysis shows that high expression of the inhibitor of apoptosis protein survivin (BIRC5) correlates with advanced stage and poor prognosis. Recent work in our lab demonstrated that survivin promotes proliferation, migration, and mitochondrial remodeling in the RENCA RCC cell model, suggesting that survivin lies at the intersection of cell cycle and metabolic control. Preliminary RNA-seq analysis of survivin siRNA-treated RENCA cells indicated that immune-recruiting cytokines are upregulated upon survivin loss, which we validated in vitro. Because RENCA does not fully recapitulate ccRCC biology, we investigated how survivin loss influences mitochondrial homeostasis and the tumor–immune interface across VHL-deficient ccRCC models. We utilized a novel murine VHL-null ccRCC line (LVRCC), the VHL-proficient RENCA line, and the human VHL-deficient 786-O ccRCC line. Survivin was depleted by siRNA, followed by assessment of mitochondrial mass and immune cell recruitment using transwell co-culture assays. Notably, survivin knockdown in LVRCC cells produced a change in mitochondrial mass opposite to that observed in RENCA, a finding potentially related to VHL status. The VHL-deficient 786-O line exhibited phenotypes similar to LVRCC cells, including survivin-dependent growth. Survivin loss increased immune cell migration toward ccRCC cells, indicating that survivin restrains immune cell infiltration into the tumor microenvironment. Collectively, these data support survivin as an oncogenic driver and putative therapeutic target in ccRCC. Citation Format: Shivani S. Tuli, Yamato Murakami, Cais Vo, Yongho Bae, David Gau. Survivin regulates tumorigenesis and immunogenicity of ccRCC [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Innovations in Kidney Cancer Research: From Molecular Insights to Therapeutic Breakthroughs; 2026 Mar 13-16; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2026;86(5_Suppl_2):Abstract nr B020.

Cardiovascular diseases (CVDs) and pathologies are often driven by changes in molecular signaling and communication, as well as in cellular and tissue components, particularly those involving the extracellular matrix (ECM), cytoskeleton, and immune response. The fine-wire vascular injury model is commonly used to study neointimal hyperplasia and vessel stiffening, but it is not typically considered a model for CVDs. However, applying this model to study CVDs in conjunction with established processes could offer valuable insights. In this paper, we hypothesize that vascular injury induces changes in gene expression, molecular communication, and biological processes similar to those observed in CVDs at both the transcriptome and protein levels. To investigate this, we analyzed gene expression in microarray datasets from injured and uninjured femoral arteries in mice two weeks post-injury, identifying 1,467 significantly and differentially expressed genes involved in several CVDs such as including vaso-occlusion, arrhythmia, and atherosclerosis. We further constructed a protein-protein interaction network with seven functionally distinct clusters, with notable enrichment in ECM, metabolic processes, actin-based process, and immune response. Significant molecular communications were observed between the clusters, most prominently among those involved in ECM and cytoskeleton organizations, inflammation, and cell cycle. Machine Learning Disease pathway analysis revealed that vascular injury-induced crosstalk between ECM remodeling and immune response clusters contributed to aortic aneurysm, neovascularization of choroid, and kidney failure. Additionally, we found that interactions between ECM and actin cytoskeletal reorganization clusters were linked to cardiac damage, carotid artery occlusion, and cardiac lesions. Overall, through multi-scale bioinformatic analyses, we demonstrated the robustness of the vascular injury model in eliciting transcriptomic and molecular network changes associated with CVDs, highlighting its potential for use in cardiovascular research.

Meniscus injuries are challenging to treat due to the tissue heterogeneity and limited treatment efficacy. Understanding meniscus cell migration, crucial for healing, remains incomplete, especially its zonal dependency. This study explores how epigenetic mechanisms affect meniscus cell migration under inflammation, focusing on healing implications. Distinct histone modifications and chromatin dynamics between inner and outer cells were observed during migration, emphasizing the need to consider these differences in repair strategies. Furthermore, tumor necrosis factor alpha (TNF-α), a proinflammatory cytokine, slows inner meniscus cell migration, while outer cells remain unaffected, indicating a zonal response. Interestingly, TNF-α differentially alters histone modifications, particularly H3K27me3, between the cell types. Transcriptome analysis showed significant gene expression changes with inner cells more affected than outer cells. Gene cluster analysis revealed different responses in chromatin remodeling, extracellular matrix assembly, and wound healing between zones. We further identified potential therapeutic targets by using epigenetic drugs, GSKJ4 (a histone demethylase inhibitor) and C646 (a histone acetyltransferase inhibitor), which restored inner meniscus cell migration under inflammatory conditions, highlighting their potential in treating meniscus tears. This highlights their potential utility in treating meniscus tear injuries. Overall, our findings elucidate the intricate interplay between epigenetic mechanisms and meniscus cell migration, along with its meniscus zonal dependency. This study provides insight into potential targets for enhancing meniscus repair and regeneration, which may lead to improved clinical outcomes for patients with meniscus injuries and osteoarthritis.

Arterial stiffness is a contributor to cardiovascular diseases (CVDs) and is associated with the aberrant migration of vascular smooth muscle cells (VSMCs). However, the mechanisms driving VSMC migration in stiff environments remain unclear. We recently demonstrated that survivin is upregulated in mouse and human VSMCs cultured on stiff hydrogels, where it modulates stiffness-mediated cell proliferation. However, its role in stiffness-dependent VSMC migration remains unknown. To assess its impact on migration, we performed time-lapse microscopy on VSMCs seeded on fibronectin-coated soft and stiff hydrogels, mimicking the physiological stiffness of normal and diseased arteries. We observed that VSMC motility increased under stiff conditions, while pharmacologic or siRNA-mediated inhibition of survivin reduced stiffness-stimulated migration to rates similar to those observed under soft conditions. Further investigation revealed that cells on stiff hydrogels exhibited greater directional movement and robust lamellipodial protrusion compared to those on soft hydrogels. Interestingly, survivin-inhibited cells on stiff hydrogels showed reduced directional persistence and lamellipodial protrusion. We also found that survivin overexpression modestly increased cell motility and partially rescued the lack of directional persistence compared to green fluorescent protein (GFP)-expressing VSMCs on soft hydrogels. Mechanistically, stiffness- and survivin-dependent cell migration involves focal adhesion kinase (FAK) and actin dynamics, as stiffness increases phosphorylated FAK recruitment to focal adhesions and promotes actin organization and stress fiber formation—effects that are disrupted by survivin inhibition. In conclusion, our findings establish that mechanotransduction through a survivin–FAK–actin cascade converts extracellular matrix stiffness into stiffness-sensitive motility, suggesting that targeting this pathway may offer therapeutic strategies for CVD.

Pulmonary fibrosis (PF) is a major cause of morbidity and mortality. Although increased oxidative stress and altered metabolism are implicated in PF pathobiology, our knowledge regarding the contribution of the glucose metabolism to the synthesis of extracellular matrix (ECM) is still incomplete. Therefore, our objective was to determine altered metabolic pathways that contribute to bleomycin (BLM; 5 mg/kg) sulfate-induced PF in rats. We determined the effects of nebulized BLM on PF in CRISPR-edited rats expressing glucose-6-phosphate dehydrogenase (G6PD) variant (S188F; G6PDS188F) and their wild-type (WT) littermates. Unexpectedly, application of BLM increased lung tissue volume in G6PDS188F rats as compared with WT littermates. Masson's Trichrome staining and Ashcroft scoring revealed increased collagen in perivascular regions and around the airways and hydroxyproline within the lungs of G6PDS188F + BLM as compared with WT + BLM rats. In addition, mass spectrometry-based proteomics and spatial proteomics confirmed increased expression of profibrotic proteins, including collagen1a1 and baculoviral IAP repeat containing 5, in the lungs of G6PDS188F + BLM rats compared with WT + BLM rats. Since BLM increased expression of KEAP1, we suggest that BLM inactivated NRF2 and increased oxidized glutathione, an indicator of oxidative stress that increases ECM, in lungs of G6PDS188F rats. Finally, unbiased metabolomics revealed downregulated spermidine, a polyamine pathway metabolite that decreases BLM-induced collagen deposition, in the lungs of G6PDS188F + BLM rats. Therefore, we propose that dysregulated polyamine pathway and antioxidant state exacerbated BLM-induced synthesis of ECM-related proteins in G6PDS188F variant rats as compared with their WT littermates.NEW & NOTEWORTHY This study reports that a loss-of-function G6PD variant exacerbates BLM-induced lung fibrosis in rats by suppressing polyamine pathway and increasing oxidative stress that oxidized the key ECM-related proteins and antioxidants.