Vascular Differentiation Research Articles

Mechanical Trapping of the Cell Nucleus Into Microgroove Concavity But Not On Convexity Induces Cell Tissue Growth and Vascular Smooth Muscle Differentiation

Mechanical Trapping of the Cell Nucleus Into Microgroove Concavity But Not On Convexity Induces Cell Tissue Growth and Vascular Smooth Muscle Differentiation

Comparative Study on Vascular Bundle Morphological Characteristics of Parts of Branches, Culms, and Rhizomes of Oligostachyum sulcatum

This study presents a comprehensive analysis of the vascular bundle morphology, tangential and radial diameters, and distribution frequency of different parts of Oligostachyum sulcatum, elucidating their structural and functional significance. Electron microscopy images revealed distinct vascular bundle characteristics in the different parts, including the vascular bundles in both parts of the rhizomes, the middle parts of the internodes, and the middle and inner parts of the branches, which were semi-open. The vascular bundles in the outer parts of both internodes and branches were semi-differentiated and undifferentiated. The vascular bundles in the inner parts of internodes were open. Statistical analysis showed significant variations in tangential and radial diameters among these parts, reflecting their diverse mechanical and physiological functions. The internodes exhibited the largest tangential and radial diameters, suggesting a critical role in mechanical support. In contrast, the branches had the smallest diameter, indicating that vascular bundle differentiation is influenced by growth conditions. The vascular bundle frequency was the highest in branches and the lowest in inside-sticks. This study provides theoretical references for the adaptive strategies and growth regulation mechanisms of O. sulcatum.

An unexpected role of IL10 in mesoderm induction and differentiation from pluripotent stem cells: Implications in zebrafish angiogenic sprouting, vascular organoid development, and therapeutic angiogenesis

Mesoderm induction is a crucial step for vascular cell specification, vascular development and vasculogenesis. However, the cellular and molecular mechanisms underlying mesoderm induction remain elusive. In the present study, a chemically-defined differentiation protocol was used to induce mesoderm formation and generate functional vascular cells including smooth muscle cells (SMCs) and endothelial cells (ECs) from human induced pluripotent stem cells (hiPSCs). Zebrafish larvae were used to detect an in vivo function of interleukin 10 (IL10) in mesoderm formation and vascular development. A three dimensional approach was used to create hiPSC-derived blood vessel organoid (BVO) and explore a potential impact of IL10 on BVO formation. A murine model hind limb ischemia was applied to investigate a therapeutic potential of hiPSC-derived cells treated with or without IL10 during differentiation. We found that IL10 was significantly and specifically up-regulated during mesoderm stage of vascular differentiation. IL10 addition in mesoderm induction media dramatically increased mesoderm induction and vascular cell generation from hiPSCs, whereas an opposite effect was observed with IL10 inhibition. Mechanistic studies revealed that IL10 promotes mesoderm formation and vascular cell differentiation by activating signal transducer and activator of transcription 3 signal pathway. Functional studies with an in vivo model system confirmed that knockdown of IL10 using morpholino antisense oligonucleotides in zebrafish larvae caused defective mesoderm formation, angiogenic sprouting and vascular development. Additionally, our data also show IL10 promotes blood vessel organoid development and enhances vasculogenesis and angiogenesis. Importantly, we demonstrate that IL10 treatment during mesoderm induction stage enhances blood flow perfusion recovery and increases vasculogenesis and therapeutic angiogenesis after hind limb ischemia. Our data, therefore, demonstrate a regulatory role for IL10 in mesoderm formation from hiPSCs and during zebrafish vascular development, providing novel insights into mesoderm induction and vascular cell specifications.

Breaking New Ground: Unraveling the USP1/ID3/E12/P21 Axis in Vascular Calcification

Vascular calcification (VC) poses significant challenges in cardiovascular health. This study employs single-cell transcriptome sequencing to dissect cellular dynamics in this process. We identify distinct cell subgroups, notably in vascular smooth muscle cells (VSMCs), and observe differences between calcified atherosclerotic cores and adjacent regions. Further exploration reveals ID3 as a key gene regulating VSMC function. In vitro experiments demonstrate ID3′s interaction with USP1 and E12, modulating cell proliferation and osteogenic differentiation. Animal models confirm the critical role of the USP1/ID3/E12/P21 axis in VC. This study sheds light on a novel regulatory mechanism, offering potential therapeutic targets.1.This study uses single-cell transcriptome to investigate vascular calcification in depth.2.This study successfully identifies cellular heterogeneity related to vascular calcification.3.This study reveals the crucial role of ID3 in vascular smooth muscle cell osteogenic differentiation.4.In vivo and in vitro experiments of this study demonstrate the regulatory role of the USP1/ID3/E12/P21 axis in calcification.5.This study provides potential molecular targets for the treatment of calcification-related diseases.

Differential regulation of xylem and phloem differentiation in grape berries by GA3 and CPPU

Fruit vascular bundles play a critical role in transporting water and nutrients within fruit, ultimately impacting fruit size and quality. Gibberellic acid (GA) and the synthetic cytokinin forchlorfenuron (CPPU) are commonly used to enhance grape berry size. Understanding their effects on the structure and function of berry vascular bundles could provide new information on the regulatory mechanism of these hormones on grapevine expansion and quality. In this study, the fruitlets of ‘Pinot Noir’ and ‘Einset Seedless’ grape varieties were treated by GA3, CPPU, and a combination of both at two weeks after full bloom. The samples were collected at 20, 40 and 70 days post-treatment. The results showed that all treatments led to an increase in berry size, xylem area, phloem area, the relative abundance of large-diameter grade vessels (>6 μm), the corresponding xylem area-specific hydraulic conductivity, and water absorption in both grape varieties. Specifically, CPPU favored an increase in phloem area, while GA3 primarily increased the xylem area. The combined application had a strong effect on both xylem and phloem area. RNA-seq analysis revealed that all treatments induced variations in the expression levels of multiple hormone signal components and genes related to vascular development. GA3 treatment mainly influenced some specific transcription factors involved in xylem differentiation, while CPPU and the combination treatment induced a wide range of genes related to vascular cambium initiation, proliferation, and differentiation of phloem and xylem vessels. Overall, our findings suggest that GA3 played a significant role in xylem differentiation, whereas CPPU had a major impact on both phloem and xylem differentiation in grape berries. These results lay the groundwork for further research the precise regulation of fruit vascular development under GA3 and CPPU treatments.

Investigating properties of sweet cherry (Prunus avium) flower buds that help promote freezing avoidance by supercooling.

Mechanisms involved in the supercooling of plant tissues as a means of low temperature survival are still not fully understood. We investigated properties that may promote supercooling in overwintering sweet cherry (Prunus avium) flower buds. We conducted experiments on sweet cherry flower buds using differential thermal analysis (DTA) and observed locations of ice formation in the bud structure. We also used anatomical development and water-soluble dye uptake throughout the overwintering period to identify changes that correlate with gain and loss of supercooling capacity. Our results revealed barriers to ice propagation are likely unique to each primordium, as inferred from exotherms produced from buds subjected to DTA, although multiple primordia may freeze simultaneously. Ice is accommodated between the bud scales and within the bud axis; however, full expression of supercooling was not dependent on the presence of scales. Anatomical and DTA studies revealed a correlation between vascular differentiation in primordia and loss of supercooling in the spring; these observations were at a higher temporal resolution than previously described for Prunus. Furthermore, disturbing tissues subtending the primordia interfered with typical patterns of supercooling, indicated more erratic and numerous exotherms produced during DTA. In summary, sweet cherry flower buds undergo extra-organ freezing. In winter, a barrier to ice propagation in the region directly subtending primordia protects the flower from freezing damage, but in the spring xylem differentiation in primordia provides a conduit for ice propagation that compromises supercooling.

The Expression of Circ-Astn1 Inhibits High Glucose Induced Endothelial Progenitor Cell Dysfunction by Activating Autophagy

ABSTRACT Background Diabetes mellitus (DM) and complications such as chronic kidney disease and cardiovascular symptoms pose a substantial public health burden. Increasing studies have shown that circular RNAs (circRNAs) regulate many gene expressions that are essential in diverse pathological and biological procedures. However, the roles of particular circRNAs in DM are unclear. Methods In the current investigation, endothelial progenitor cells (EPCs) were used to search for abnormal expression of circRNAs by using high-throughput sequencing under high glucose (HG) conditions. The regulatory mechanisms and targets were then studied through bioinformatics analysis, luciferase reporter analysis, angiogenic differentiation experiments, flow cytometry detection of apoptosis and RT-qPCR analysis. Results The circ-Astn1 expression in EPCs decreased after HG treatment. Overexpression or circ-Astn1 suppressed HG induced endothelial cell damage. MicroRNA (miR)-138-5p and SIRT5 were found to be the downstream targets of circ-Astn1 through luciferase reporter analysis. SIRT5 downregulation or miR-138-5p overexpression reversed circ-Astn1’s protective effect against HG induced endothelial cell dysfunction, including apoptosis and abnormal vascular differentiation. Furthermore, circ-Astn1 overexpression promoted autophagy activation by increasing SIRT5 expression under HG conditions. Our findings suggest that circ-Astn1 mediated promotion of SIRT5 facilitates autophagy by sponging miR-138-5p. Conlusion Together, our findings show that the overexpression of circ-Astn1 suppresses HG induced endothelial cell damage by targeting miR-138-5p/SIRT5 axis.

Placenta‐Derived Mesenchymal Stromal‐Like Cells Promote 3D‐Engineered Muscle Tissue Differentiation and Vessel Network Maturation

Placental‐derived stromal‐like cells (PLX‐PAD) have been shown to facilitate muscle tissue recovery after injury and stimulate angiogenesis. This work assesses the impact of PLX‐PAD cells on the vascularization and maturation of engineered skeletal muscle tissue. Specifically, their effects in direct co‐culture with endothelial cells, pericytes, and myoblasts seeded within microporous 3D scaffolds are characterized. Additionally, the impact of hypoxic PLX‐PAD cell‐conditioned medium (CM) on vascularization and muscle differentiation of engineered tissue is monitored. Co‐culture of PLX‐PAD with myocytes stimulated myocyte differentiation while PLX‐PAD CM promoted the formation of vascular networks. Implantation of a multi‐culture system of vascularized human skeletal muscle tissue and PLX‐PAD into a rectus abdominal defect in nude mice promoted myocyte differentiation, host vessel penetration, and tissue integration. These findings indicate the ability of placenta‐derived cells to induce the formation of vascularized engineered muscle constructs with potential therapeutic applications.

Photothermal switch by gallic acid-calcium grafts synthesized by coordination chemistry for sequential treatment of bone tumor and regeneration

The residual bone tumor and defects which is caused by surgical therapy of bone tumor is a major and important problem in clinicals. And the sequential treatment for irradiating residual tumor and repairing bone defects has wildly prospects. In this study, we developed a general modification strategy by gallic acid (GA)-assisted coordination chemistry to prepare black calcium-based materials, which combines the sequential photothermal therapy of bone tumor and bone defects. The GA modification endows the materials remarkable photothermal properties. Under the near-infrared (NIR) irradiation with different power densities, the black GA-modified bone matrix (GBM) did not merely display an excellent performance in eliminating bone tumor with high temperature, but showed a facile effect of the mild-heat stimulation to accelerate bone regeneration. GBM can efficiently regulate the microenvironments of bone regeneration in a spatial-temporal manner, including inflammation/immune response, vascularization and osteogenic differentiation. Meanwhile, the integrin/PI3K/Akt signaling pathway of bone marrow mesenchymal stem cells (BMSCs) was revealed to be involved in the effect of osteogenesis induced by the mild-heat stimulation. The outcome of this study not only provides a serial of new multifunctional biomaterials, but also demonstrates a general strategy for designing novel blacked calcium-based biomaterials with great potential for clinical use.

Methylome analysis of endothelial cells suggests new insights on sporadic brain arteriovenous malformation

Arteriovenous malformation of the brain (bAVM) is a vascular phenotype related to brain defective angiogenesis. Involved vessels show impaired expression of vascular differentiation markers resulting in the arteriolar to venule direct shunt. In order to clarify aberrant gene expression occurring in bAVM, here we describe results obtained by methylome analysis performed on endothelial cells (ECs) isolated from bAVM specimens, compared to human cerebral microvascular ECs. Results were validated by quantitative methylation-specific PCR and quantitative realtime-PCR. Differential methylation events occur in genes already linked to bAVM onset, as RBPJ and KRAS. However, among differentially methylated genes, we identified EPHB1 and several other loci involved in EC adhesion as well as in EC/vascular smooth muscle cell (VSMC) crosstalk, suggesting that only endothelial dysfunction might not be sufficient to trigger the bAVM phenotype. Moreover, aberrant methylation pattern was reported for many lncRNA genes targeting transcription factors expressed during neurovascular development. Among these, the YBX1 that was recently shown to target the arteridin coding gene. Finally, in addition to the conventional CpG methylation, we further considered the role of impaired CHG methylation, mainly occurring in brain at embryo stage. We showed as differentially CHG methylated genes are clustered in pathways related to EC homeostasis, as well as to VSMC-EC crosstalk, suggesting as impairment of this interaction plays a prominent role in loss of vascular differentiation, in bAVM phenotype.

The REPLUMLESS Transcription Factor Controls the Expression of the RECEPTOR-LIKE CYTOPLASMIC KINASE VI_A2 Gene Involved in Shoot and Fruit Patterning of Arabidopsis thaliana.

The promoter of the RECEPTOR-LIKE CYTOPLASMIC KINASE VI_A2 (RLCK VI_A2) gene contains nine binding sites for the REPLUMLESS (RPL) transcription factor. In agreement, the expression of the kinase gene was strongly downregulated in the rpl-4 mutant. Comparing phenotypes of loss-of-function mutants, it was revealed that both genes are involved in stem growth, phyllotaxis, organization of the vascular tissues, and the replum, highlighting potential functional interactions. The expression of the RLCKVI_A2 gene from the constitutive 35S promoter could not complement the rpl-4 phenotypes but exhibited a dominant positive effect on stem growth and affected vascular differentiation and organization. The results also indicated that the number of vascular bundles is regulated independently from stem thickness. Although our study cannot demonstrate a direct link between the RPL and RLVKVI_A2 genes, it highlights the significance of the proper developmental regulation of the RLCKVI_A2 promoter for balanced stem development.

3D-printed bone regeneration scaffolds modulate bone metabolic homeostasis through vascularization for osteoporotic bone defects

The treatment of osteoporotic bone defects poses a challenge due to the degradation of the skeletal vascular system and the disruption of local bone metabolism within the osteoporotic microenvironment. However, it is feasible to modulate the disrupted local bone metabolism imbalance through enhanced vascularization, a theory termed "vascularization-bone metabolic balance". This study developed a 3D-printed polycaprolactone (PCL) scaffold modified with EPLQLKM and SVVYGLR peptides (PCL-SE). The EPLQLKM peptide attracts bone marrow-derived mesenchymal stem cells (BMSCs), while the SVVYGLR peptide enhances endothelial progenitor cells (EPCs) vascular differentiation, thus regulating bone metabolism and fostering bone regeneration through the paracrine effects of EPCs. Further mechanistic research demonstrated that PCL-SE promoted the vascularization of EPCs, activating the Notch signaling pathway in BMSCs, leading to the upregulation of osteogenesis-related genes and the downregulation of osteoclast-related genes, thereby restoring bone metabolic balance. Furthermore, PCL-SE facilitated the differentiation of EPCs into "H"-type vessels and the recruitment of BMSCs to synergistically enhance osteogenesis, resulting in the regeneration of normal microvessels and bone tissues in cases of femoral condylar bone defects in osteoporotic SD rats. This study suggests that PCL-SE supports in-situ vascularization, remodels bone metabolic translational balance, and offers a promising therapeutic regimen for osteoporotic bone defects.

Internal flow field analysis of a dendritic pore scaffold for bone tissue engineering

The effective reconstruction of osteochondral biomimetic structures is a key factor in guiding the regeneration of full-thickness osteochondral defects. Due to the avascular nature of hyaline cartilage, the greatest challenge in constructing this scaffold lies in both utilizing the biomimetic structure to promote vascular differentiation for nutrient delivery to hyaline cartilage, thereby enhancing the efficiency of osteochondral reconstruction, and effectively blocking vascular ingrowth into the cartilage layer to prevent cartilage mineralization. However, the intrinsic relationship between the planning of the microporous pipe network and the flow resistance in the biomimetic structure, and the mechanism of promoting cell adhesion to achieve vascular differentiation and inhibiting cell adhesion to block the growth of blood vessels are still unclear. Inspired by the structure of tree trunks, this study designed a biomimetic tree-like tubular network structure for osteochondral scaffolds based on Murray’s law. Utilizing computational fluid dynamics, the study investigated the influence of the branching angle of micro-pores on the flow velocity, pressure distribution, and scaffold permeability within the scaffold. The results indicate that when the differentiation angle exceeds 50 degrees, the highest flow velocity occurs at the confluence of tributaries at the ninth fractal position, forming a barrier layer. This structure effectively guides vascular growth, enhances nutrient transport capacity, increases flow velocity to promote cell adhesion, and inhibits cell infiltration into the cartilage layer.

YABBY and diverged KNOX1 genes shape nodes and internodes in the stem.

Plant stems comprise nodes and internodes that specialize in solute exchange and elongation. However, their boundaries are not well defined, and how these basic units arise remains elusive. In rice with clear nodes and internodes, we found that one subclade of class I knotted1-like homeobox (KNOX1) genes for shoot meristem indeterminacy restricts node differentiation and allows internode formation by repressing YABBY genes for leaf development and genes from another node-specific KNOX1 subclade. YABBYs promote nodal vascular differentiation and limit stem elongation. YABBY and node-specific KNOX1 genes specify the pulvinus, which further elaborates the nodal structure for gravitropism. Notably, this KNOX1 subclade organization is specific to seed plants. We propose that nodes and internodes are distinct domains specified by YABBY-KNOX1 cross-regulation that diverged in early seed plants.

Endothelial cell Piezo1 promotes vascular smooth muscle cell differentiation on large arteries.

Vascular stabilization is a mechanosensitive process, in part driven by blood flow. Here, we demonstrate the involvement of the mechanosensitive ion channel, Piezo1, in promoting arterial accumulation of vascular smooth muscle cells (vSMCs) during zebrafish development. Using a series of small molecule antagonists or agonists to temporally regulate Piezo1 activity, we identified a role for the Piezo1 channel in regulating klf2a levels and altered targeting of vSMCs between arteries and veins. Increasing Piezo1 activity suppressed klf2a and increased vSMC association with the cardinal vein, while inhibition of Piezo1 activity increased klf2a levels and decreased vSMC association with arteries. We supported the small molecule data with in vivo genetic suppression of piezo1 and 2 in zebrafish, resulting in loss of transgelin+ vSMCs on the dorsal aorta. Further, endothelial cell (EC)-specific Piezo1 knockout in mice was sufficient to decrease vSMC accumulation along the descending dorsal aorta during development, thus phenocopying our zebrafish data, and supporting functional conservation of Piezo1 in mammals. To determine mechanism, we used in vitro modeling assays to demonstrate that differential sensing of pulsatile versus laminar flow forces across endothelial cells changes the expression of mural cell differentiation genes. Together, our findings suggest a crucial role for EC Piezo1 in sensing force within large arteries to mediate mural cell differentiation and stabilization of the arterial vasculature.

Modulation of FGF pathway signaling and vascular differentiation using designed oligomeric assemblies

Many growth factors and cytokines signal by binding to the extracellular domains of their receptors and driving association and transphosphorylation of the receptor intracellular tyrosine kinase domains, initiating downstream signaling cascades. To enable systematic exploration of how receptor valency and geometry affect signaling outcomes, we designed cyclic homo-oligomers with up to 8 subunits using repeat protein building blocks that can be modularly extended. By incorporating a de novo-designed fibroblast growth factor receptor (FGFR)-binding module into these scaffolds, we generated a series of synthetic signaling ligands that exhibit potent valency- and geometry-dependent Ca2+ release and mitogen-activated protein kinase (MAPK) pathway activation. The high specificity of the designed agonists reveals distinct roles for two FGFR splice variants in driving arterial endothelium and perivascular cell fates during early vascular development. Our designed modular assemblies should be broadly useful for unraveling the complexities of signaling in key developmental transitions and for developing future therapeutic applications.

A promising approach for quantifying focal stroke modeling and assessing stroke progression: optical resolution photoacoustic microscopy photothrombosis.

JOURNAL/nrgr/04.03/01300535-202507000-00025/figure1/v/2024-09-09T124005Z/r/image-tiff To investigate the mechanisms underlying the onset and progression of ischemic stroke, some methods have been proposed that can simultaneously monitor and create embolisms in the animal cerebral cortex. However, these methods often require complex systems and the effect of age on cerebral embolism has not been adequately studied, although ischemic stroke is strongly age-related. In this study, we propose an optical-resolution photoacoustic microscopy-based visualized photothrombosis methodology to create and monitor ischemic stroke in mice simultaneously using a 532 nm pulsed laser. We observed the molding process in mice of different ages and presented age-dependent vascular embolism differentiation. Moreover, we integrated optical coherence tomography angiography to investigate age-associated trends in cerebrovascular variability following a stroke. Our imaging data and quantitative analyses underscore the differential cerebrovascular responses to stroke in mice of different ages, thereby highlighting the technique's potential for evaluating cerebrovascular health and unraveling age-related mechanisms involved in ischemic strokes.

Genome wide characterization and expression insights of Auxin Response Factor (ARF) gene family in cabbage in response to clubroot disease

Auxin response factors (ARFs) regulate genes expression in response to auxin. Auxin performs a significant role in plant development and growth such as vascular tissue differentiation, root initiation and apical dominance. Within plants, auxin promotes the growth of vascular tissue differentiation, apical dominance and at the cellular level, auxin helps cells to divide and differentiate. Members of the ARF family carry a DNA binding domain (DBD) that binds to specific DNA sequences in the promoters of target genes. Despite the crucial role of the ARF gene family, no comprehensive study has been carried out to characterize the comparative ARF in B oleracea and A thaliana. For this reason, an in-silico approach was adopted. During the study, we found 49 genes in B oleracea and 37 genes in A thaliana. Of these, only 17 and 22 genes were studied due to their redundancy. In the phylogenetic analysis, we included B oleracea, B rapa, B napus and A thaliana and divided them into four groups based on conserved domains. Chromosomal mapping identified the positions of the genes on different chromosomes. The diversity of introns in the gene structure varies from 1 to 13 and up to five protein motifs are shown on each gene. Many transcription factors were identified by cis-regulatory elements. Gene expression revealed the involvement of BolARFs in root gall disease. The presence of nine different conserved domains was observed, of which B3, AUX_IAA superfamily together with auxin response were dominantly repeated in maximum number of genes. The overall study provides new insights and directions for further research and analysis in the future.

Auxin signaling in the cambium promotes tissue adhesion and vascular formation during Arabidopsis graft healing.

The strong ability of plants to regenerate wounds is exemplified by grafting when two plants are cut and joined together to grow as one. During graft healing, tissues attach, cells proliferate, and the vasculatures connect to form a graft union. The plant hormone auxin plays a central role, and auxin-related mutants perturb grafting success. Here, we investigated the role of individual cell types and their response to auxin during Arabidopsis (Arabidopsis thaliana) graft formation. By employing a cell-specific inducible misexpression system, we blocked auxin response in individual cell types using the bodenlos mutation. We found that auxin signaling in procambial tissues was critical for successful tissue attachment and vascular differentiation. In addition, we found that auxin signaling was required for cell divisions of the procambial cells during graft formation. Loss of function mutants in cambial pathways also perturbed attachment and phloem reconnection. We propose that cambial and procambial tissues drive tissue attachment and vascular differentiation during successful grafting. Our study thus refines our knowledge of graft development and furthers our understanding of the regenerative role of the cambium.

Chronic Gestational Hypoxia Alters Plasma Exosomal miRNA Profile and Pulmonary Arterial Proteome and Metabolome in Fetal Lambs

Antenatal chronic hypoxia negatively impacts fetal development and increases the risk of cardio-pulmonary dysfunction after birth, including newborn pulmonary hypertension. Previously, we showed that fetal and newborn sheep exposed to chronic hypoxia during gestation exhibit a variety of cardio-pulmonary defects that parallel those of human infants including pulmonary vascular remodeling, heart dysfunction, and pulmonary hypertension. To explore the mechanisms involved in hypoxia-mediated newborn pulmonary hypertension, we test the hypothesis that antenatal chronic hypoxia impairs pathways critical to pulmonary vascular development that are coupled to abnormalities in lung structure and function through dysregulation in cell growth and proliferation. To address this hypothesis, we employed an animal model of pregnant sheep exposed to high altitude hypoxia at the White Mountain Research Station, which has an altitude of 3801 meters. Hypoxic pregnant sheep were housed at altitude during gestation for ~110 days out of 138-141 days of pregnancy while normoxic counterparts were kept at 700 meters. Animals were transferred to Loma Linda at 355 meters and fetal lambs studied at ~140 days of gestation. Fetal plasma samples were collected for exosomal miRNA analysis and pulmonary arteries were collected for proteomic and metabolomic analysis. Ingenuity pathway analysis and other bioinformatic tools were used to interrogate regulatory mechanisms and pathways and the web of molecular interactions affected by hypoxia. Antenatal hypoxic stress caused up and down regulation of a limited number of plasma exosomal miRNAs, pulmonary arterial metabolites and proteins, with discrete interconnections between the various analytes and molecules. There were marked changes in upstream regulators and markers that point to increased oxidative stress and inflammatory response along with pronounced downregulation of matrix proteins and phenotypic markers of vascular smooth muscle cell differentiation. Pathway analyses indicate antenatal chronic hypoxia dysregulates growth and proliferation of vascular cells and impinges on cell migration. While these data require additional study and verification, they provide new insights in the understanding of mechanisms underlying the development of pulmonary hypertension in the newborn following antenatal hypoxia. This work was supported by the National Institutes of Health Grants NIH R01HL155295, R01HL149608, R03HD098477, P01HD083132, and U24DK097154. Additional support was provided by the Loma Linda University School of Medicine, A pilot Project through the West Coast Metabolomics Center, the University of Redlands Summer Research Program, and the Walter E. Macpherson Society. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.