Biology subjects

Pena, B.

Publications and source records attributed to Pena, B..

7 recordsLinked to original sources

bioRxiv2026

Translating Sex- and Disease-Informed Design Criteria from Human Lung into Engineered Biomaterials

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease characterized by pathological extracellular matrix (ECM) remodeling, and a pronounced sex disparity, with approximately 70% of cases occurring in males. Despite these differences, biomaterials used to model fibrosis are commonly designed to replicate disease state or stiffness alone and rarely account for biological sex. Here, we derived sex- and disease-informed biomaterial design criteria from female and male control and IPF human lungs and translated these criteria into engineered hybrid-hydrogels containing human lung decellularized ECM (dECM) and poly(ethylene glycol) alpha methacrylate (PEGMA). Matrisome-focused reanalysis of our previously published region-resolved human lung proteomic dataset revealed region- and sex-associated differences in fibrillar collagens, matricellular proteins, and structural proteoglycans. Complementary characterization of human lung tissue further identified sex- and disease-associated differences in fibrillar collagen architecture, proteoglycan and glycosaminoglycan (GAG) remodeling, and tissue mechanics. Male IPF lungs exhibited the highest levels of disease-associated stiffening and fibrillar collagen remodeling, while female IPF tissue showed comparatively modest mechanical changes despite biochemical remodeling. These measurements established tissue-informed design criteria spanning matrix composition, architecture, and mechanics. We next evaluated whether tissue-derived properties were reproduced or retained following decellularization, processing, and incorporation of donor-derived dECM into hybrid-hydrogels. Engineered hybrid-hydrogels closely recapitulated sex- and disease-associated mechanical relationships, while COL1A1-associated abundance patterns were more consistently preserved than fibrillar collagen organization. Matricellular proteins and GAG-rich matrix remained detectable, although condition-dependent biochemical fidelity varied. This work establishes a human tissue-informed framework for engineering sex- and disease-specific biomaterials, highlighting the importance of defining sex-associated ECM properties and validating individual features following material fabrication to advance human models of IPF and other diseases with sex-associated differences in susceptibility and progression.

bioengineering↗

bioRxiv2026

Laminin-511 protects pancreatic β-cells from cytokine-induced death through integrin-mediated pro-survival signaling and modulation of protein kinase C δ

During the progression of type 1 diabetes (T1D), the extracellular matrix (ECM) surrounding pancreatic islets is degraded concurrent with infiltration of autoreactive immune cells and {beta}-cell death. Among the lost ECM proteins, laminin-511 is known to be essential for islet survival under healthy and T1D associated conditions, including high levels of pro-inflammatory cytokines. However, the key {beta}-cell signaling pathways regulated by laminin and the contributions to T1D pathogenesis when these cues are lost are poorly understood. This study utilizes a biomimetic reverse thermal gel (RTG) with laminin-511 to determine if laminin protects {beta}-cells against cytokine-induced death and elucidate the signaling pathways involved. MIN6 cells, C57Bl/6 mouse islets and human islets were encapsulated in RTG scaffolds with laminin-511 and treated with a cytokine cocktail for 24 hours. Islet viability and the activities of several pro- and anti-apoptotic proteins were studied. Laminin-511 was shown to protect islets against cytokine-induced death by interacting with {beta}1 integrins and activating pro-survival Akt signaling. Pro-survival signaling was mediated by reduced activity of protein kinase C{delta} (PKC{delta}), a key mediator of cytokine-induced {beta}-cell death, at the cell membrane in the presence of laminin via reduced levels of diacylglycerol (DAG), a canonical activator of PKC{delta}. Taken together, these results demonstrate that laminin-511 is an essential factor in protecting {beta}-cells against cytokine-induced death by downregulation of membrane DAG, inhibiting activation of pro-apoptotic PKC{delta}. Our results suggest that loss of ECM in T1D may make {beta}-cells more susceptible to cytokine-induced death by increasing activation of PKC{delta}. HighlightsO_LILaminin-511 improves islet survival under cytokine treatment C_LIO_LIPro-survival Akt is upregulated by laminin-511 C_LIO_LIPro-apoptotic PKC{delta} activity is downregulated at the cell membrane by laminin-511 C_LIO_LILaminin-511 decreases membrane DAG levels leading to reduced PKC{delta} activation C_LIO_LILaminin-511 is an essential ECM component for islet survival during type 1 diabetes C_LI

cell biology↗

bioRxiv2025

A novel Notch and WNT signaling mechanism contribute to pediatric DCM: a pathway to new therapeutics.

BackgroundTherapies for pediatric idiopathic dilated cardiomyopathy (iDCM) are extrapolated from adult heart failure despite limited efficacy, suggesting fundamental biological differences. Our prior transcriptomic studies indicate activation of developmental signaling pathways, including Notch and WNT, in pediatric iDCM; however, their mechanistic contribution remains unknown. We tested whether reactivation of Notch and WNT/{beta}-catenin signaling drives pathological remodeling in postnatal hearts and whether pathway inhibition improves cardiac function. MethodsWe developed a juvenile rat model to reproduce age-dependent molecular features of pediatric iDCM using {beta}-adrenergic stimulation (isoproterenol, ISO) and secreted frizzled-related protein-1 (sFRP1), a circulating WNT modulator elevated in children with DCM. Cardiac function was assessed by echocardiography; pathway activation by immunoblotting and transcriptomics; myocardial stiffness by atomic force microscopy. Findings were compared with explanted pediatric and adult human myocardium. ResultsExplanted pediatric, but not adult iDCM hearts exhibited increased nuclear and cytoplasmic Notch intracellular domain (NICD) and {beta}-catenin. Combined ISO and sFRP1 treatment recapitulated key features of pediatric disease, including ventricular dilation, reduced ejection fraction, reactivation of the fetal gene program, and increased myocardial stiffness in the absence of fibrosis or hypertrophy. Bulk and single-nucleus RNA sequencing identified cardiomyocyte-specific activation of Notch and WNT pathways and reduced intercellular signaling diversity. Mechanistically, {beta}-catenin silencing attenuated Notch target gene activation and pathological remodeling in vitro. Pharmacologic Notch inhibition reduced NICD and {beta}-catenin accumulation, improved ventricular function, and normalized myocardial stiffness in vivo. ConclusionPediatric iDCM is characterized by pathological co-activation of developmental Notch-WNT signaling pathways that are not observed in adult disease. Reactivation of this axis promotes maladaptive remodeling and myocardial stiffening, and its inhibition improves cardiac function. These findings establish developmental signaling reactivation as a central mechanism of pediatric iDCM and support age-specific therapeutic strategies.

molecular biology↗

bioRxiv2024

The pericardium forms as a distinct structure during heart formation

The heart integrates diverse cell lineages into a functional unit, including the pericardium, a mesothelial sac that supports heart movement, homeostasis, and immune responses. However, despite its critical roles, the developmental origins of the pericardium remain uncertain due to disparate models. Here, using live imaging, lineage tracking, and single-cell transcriptomics in zebrafish, we find the pericardium forms within the lateral plate mesoderm from dedicated anterior mesothelial progenitors and distinct from the classic heart field. Imaging of transgenic reporters in zebrafish documents lateral plate mesoderm cells that emerge lateral of the classic heart field and among a continuous mesothelial progenitor field. Single-cell transcriptomics and trajectories of hand2-expressing lateral plate mesoderm reveal distinct populations of mesothelial and cardiac precursors, including pericardial precursors that are distinct from the cardiomyocyte lineage. The mesothelial gene expression signature is conserved in mammals and carries over to postnatal development. Light sheet-based live-imaging and machine learning-supported cell tracking documents that during heart tube formation, pericardial precursors that reside at the anterior edge of the heart field migrate anteriorly and medially before fusing, enclosing the embryonic heart to form a single pericardial cavity. Pericardium formation proceeds even upon genetic disruption of heart tube formation, uncoupling the two structures. Canonical Wnt/{beta}-catenin signaling modulates pericardial cell number, resulting in a stretched pericardial epithelium with reduced cell number upon canonical Wnt inhibition. We connect the pathological expression of secreted Wnt antagonists of the SFRP family found in pediatric dilated cardiomyopathy to increased pericardial stiffness: sFRP1 in the presence of increased catecholamines causes cardiomyocyte stiffness in neonatal rats as measured by atomic force microscopy. Altogether, our data integrate pericardium formation as an independent process into heart morphogenesis and connect disrupted pericardial tissue properties such as pericardial stiffness to pediatric cardiomyopathies.

developmental biology↗

bioRxiv2024

Y chromosome linked UTY modulates sex differences in valvular fibroblast methylation in response to nanoscale extracellular matrix cues

Aortic valve stenosis (AVS) is a progressive disease wherein males more often develop valve calcification relative to females that develop valve fibrosis. Valvular interstitial cells (VICs) aberrantly activate to myofibroblasts during AVS, driving the fibrotic valve phenotype in females. Myofibroblasts further differentiate into osteoblast-like cells and produce calcium nanoparticles, driving valve calcification in males. We hypothesized the lysine demethylase UTY (ubiquitously transcribed tetratricopeptide repeat containing, Y-linked) decreases methylation uniquely in male VICs responding to nanoscale extracellular matrix cues to promote an osteoblast-like cell phenotype. Here, we describe a hydrogel biomaterial cell culture platform to interrogate how nanoscale cues modulate sex-specific methylation states in VICs activating to myofibroblasts and osteoblast-like cells. We found UTY modulates the osteoblast-like cell phenotype in response to nanoscale cues uniquely in male VICs. Overall, we reveal a novel role of UTY in the regulation of calcification processes in males during AVS progression.

bioengineering↗

bioRxiv2023

Tissue-engineered models of lung cancer premalignancy

Lung cancer is the leading global cause of cancer-related deaths. Although smoking cessation is the best preventive action, nearly 50% of all lung cancer diagnoses occur in people who have already quit smoking. Research into treatment options for these high-risk patients has been constrained to rodent models of chemical carcinogenesis, which are time-consuming, expensive, and require large numbers of animals. Here we show that embedding precision-cut lung slices within an engineered hydrogel and exposing this tissue to a carcinogen from cigarette smoke creates an in vitro model of lung cancer premalignancy. Hydrogel formulations were selected to promote early lung cancer cellular phenotypes and extend PCLS viability up to six weeks. In this study, hydrogel-embedded lung slices were exposed to the cigarette smoke derived carcinogen vinyl carbamate, which induces adenocarcinoma in mice. At six weeks, analysis of proliferation, gene expression, histology, tissue stiffness, and cellular content revealed that vinyl carbamate induced the formation of premalignant lesions with a mixed adenoma/squamous phenotype. Two putative chemoprevention agents were able to freely diffuse through the hydrogel and induce tissue-level changes. The design parameters selected using murine tissue were validated with hydrogel-embedded human PCLS and results showed increased proliferation and premalignant lesion gene expression patterns. This tissue-engineered model of human lung cancer premalignancy is the starting point for more sophisticated ex vivo models and a foundation for the study of carcinogenesis and chemoprevention strategies.

bioengineering↗

bioRxiv2022

3D-bioprinted, phototunable hydrogel models for studying adventitial fibroblast activation in pulmonary arterial hypertension

Pulmonary arterial hypertension (PAH) is a progressive disease of the lung vasculature, characterized by elevated pulmonary blood pressure, remodeling of the pulmonary arteries, and ultimately right ventricular failure. Therapeutic interventions for PAH are limited in part by the lack of in vitro screening platforms that accurately reproduce dynamic arterial wall mechanical properties. Here we present a 3D-bioprinted model of the pulmonary arterial adventitia comprised of a phototunable poly(ethylene glycol) alpha methacrylate (PEG-MA)-based hydrogel and primary human pulmonary artery adventitia fibroblasts (HPAAFs). This unique biomaterial emulates PAH pathogenesis in vitro through a two-step polymerization reaction. First, PEG-MA macromer was crosslinked off-stoichiometry by 3D bioprinting an acidic bioink solution into a basic gelatin support bath initiating a base-catalyzed thiol-ene reaction with synthetic and biodegradable crosslinkers. Then, matrix stiffening was induced by photoinitiated homopolymerization of unreacted MA end groups. A design of experiments approach produced a hydrogel platform that exhibited an initial elastic modulus (E) within the range of healthy pulmonary arterial tissue (E = 4.7 {+/-} 0.09 kPa) that was stiffened to the pathologic range of hypertensive tissue (E = 12.8 {+/-} 0.47 kPa) and supported cellular proliferation over time. A higher percentage of HPAAFs cultured in stiffened hydrogels expressed the fibrotic marker alpha-smooth muscle actin than cells in soft hydrogels (88 {+/-} 2% versus 65 {+/-} 4%). Likewise, a greater percentage of HPAAFs were positive for the proliferation marker 5-ethynyl-2-deoxyuridine (EdU) in stiffened models (66 {+/-} 6%) compared to soft (39 {+/-} 6%). These results demonstrate that 3D-bioprinted, phototunable models of pulmonary artery adventitia are a tool that enable investigation of fibrotic pathogenesis in vitro.

bioengineering↗