# Plasma Extracellular Vesicles from Bronchopulmonary Dysplasia Infants Initiate Inflammation and Abnormal Angiogenesis in Neonatal Murine Retinas **Authors:** Huijun Yuan, Matthew R. Duncan, Shaoyi Chen, Merline Benny, Augusto Schmidt, Karen Young, Audina M. Berrocal, M. Elizabeth Hartnett, Shu Wu PMC · DOI: 10.3390/cells15040367 · 2026-02-19 ## TL;DR Extracellular vesicles from infants with bronchopulmonary dysplasia cause inflammation and abnormal blood vessel growth in mice retinas, suggesting a link to retinopathy of prematurity. ## Contribution This study identifies extracellular vesicles from BPD infants as a novel contributor to retinal inflammation and abnormal angiogenesis in neonatal models. ## Key findings - BPD-EVs induce activated retinal microglia and Müller cells in neonatal mice. - BPD-EVs cause abnormal neovascularization in mouse retinas compared to nBPD-EVs. - BPD-EVs have elevated levels of inflammation and angiogenesis-related proteins. ## Abstract What are the main findings? Adoptive transfer of plasma extracellular vesicles from bronchopulmonary dysplasia infants (BPD-EVs) into neonatal mice on postnatal day 3 (P3) induced activated retinal microglia, Müller cells, and abnormal neovascularization on P17 compared to nBPD-EVs.Proteomics analysis of BPD-EVs and nBPD-EVs revealed that BPD-EVs had significantly elevated levels of inflammation and angiogenesis-related proteins compared to nBPD-EVs. Adoptive transfer of plasma extracellular vesicles from bronchopulmonary dysplasia infants (BPD-EVs) into neonatal mice on postnatal day 3 (P3) induced activated retinal microglia, Müller cells, and abnormal neovascularization on P17 compared to nBPD-EVs. Proteomics analysis of BPD-EVs and nBPD-EVs revealed that BPD-EVs had significantly elevated levels of inflammation and angiogenesis-related proteins compared to nBPD-EVs. What are the implications of the main findings? Circulating EVs may contribute to retinopathy of prematurity (ROP) development in BPD infants by inducing retinal inflammasome-associated inflammation and abnormal angiogenesis.Inflammation and angiogenesis-related proteins in BPD-EVs may represent novel biomarkers and therapeutic targets for the treatment of ROP. Circulating EVs may contribute to retinopathy of prematurity (ROP) development in BPD infants by inducing retinal inflammasome-associated inflammation and abnormal angiogenesis. Inflammation and angiogenesis-related proteins in BPD-EVs may represent novel biomarkers and therapeutic targets for the treatment of ROP. Purpose: To investigate the mechanisms by which plasma extracellular vesicles (EVs) from preterm infants with bronchopulmonary dysplasia (BPD) elicit inflammation and abnormal angiogenesis in neonatal mouse retinas. Methods: EVs from the plasma of 7-day-old preterm infants, born between 230/7 and 296/7 weeks of gestation, with BPD or without BPD (nBPD) at 36 weeks postmenstrual ages, were adoptively transferred into postnatal day 3 (P3) mice via intravenous retro-orbital sinus injection. Inflammation and pathological neovascularization in neonatal mouse retinas were examined by immunohistochemistry of retinal flat mounts for Allograft Inflammatory Factor 1 (AIF1), CD206, or Glial Fibrillary Acidic Protein (GFAP) and isolectin-B4 (IB4) staining on P17. Retinal inflammation-related transcripts were assessed by qRT-PCR. Proteomic profiles of BPD and nBPD EVs were examined by Liquid Chromatograph Mass Spectrometer/Mass Spectrometer (LC-MS/MS) and Gene Set Enrichment Analysis (GSEA). Results: Adoptively transferred EVs from BPD and nBPD infants crossed the blood–retinal barrier (BRB) in recipient mouse pups. BPD-EVs increased retinal activated microglia, Müller cells, and twisted proliferative neovascularization compared to nBPD-EVs. BPD-EVs also elevated retinal transcripts regulating inflammation and angiogenesis, including NOD-, LRR- and pyrin domain-containing protein 3 (Nlrp3), Apoptosis-associated speck-like protein containing a caspase recruitment domain (Asc), Caspase 3 (Casp3), Caspase 8 (Casp8), Gasdermin D (Gsdmd), Il1β, Il6, Aif1, and Vascular endothelial growth factor (Vegf). Proteomics analysis revealed that BPD-EVs had significantly elevated levels of inflammation and angiogenesis-related proteins compared to nBPD-EVs. Conclusions: BPD-EVs promote inflammation and abnormal neovascularization by upregulating genes related to apoptosis and inflammation in neonatal mouse retinas. EV protein profiles suggest that elevated levels of proteins such as Defensin alpha 1B (DEFA1B), Insulin-like growth factor binding protein 2 (IGFBP2), CD5 antigen-like (CD5L), von Willebrand factor (vWF), and Tenascin C (TNC) in BPD-EVs may contribute to the observed inflammation and angiogenesis. ## Linked entities - **Genes:** AIF1 (allograft inflammatory factor 1) [NCBI Gene 199], MRC1 (mannose receptor C-type 1) [NCBI Gene 4360], GFAP (glial fibrillary acidic protein) [NCBI Gene 2670], NLRP3 (NLR family pyrin domain containing 3) [NCBI Gene 114548], STS (steroid sulfatase) [NCBI Gene 412], CASP3 (caspase 3) [NCBI Gene 836], CASP8 (caspase 8) [NCBI Gene 841], GSDMD (gasdermin D) [NCBI Gene 79792], IL1B (interleukin 1 beta) [NCBI Gene 3553], IL6 (interleukin 6) [NCBI Gene 3569], VEGFA (vascular endothelial growth factor A) [NCBI Gene 7422], DEFA1B (defensin alpha 1B) [NCBI Gene 728358], IGFBP2 (insulin like growth factor binding protein 2) [NCBI Gene 3485], CD5L (CD5 molecule like) [NCBI Gene 922], VWF (von Willebrand factor) [NCBI Gene 7450], TNC (tenascin C) [NCBI Gene 3371] - **Diseases:** bronchopulmonary dysplasia (MONDO:0019091), retinopathy of prematurity (MONDO:0006952) - **Species:** Mus musculus (taxon 10090) ## Full-text entities - **Genes:** VWF (von Willebrand factor) [NCBI Gene 7450] {aka F8VWF, VWD}, Cd81 (CD81 antigen) [NCBI Gene 12520] {aka Tapa-1, Tapa1, Tspan28}, Mrc1 (mannose receptor, C type 1) [NCBI Gene 17533] {aka CD206, MR}, Pycard (PYD and CARD domain containing) [NCBI Gene 66824] {aka 9130417A21Rik, Asc, CARD5, TMS-1, TNS1, masc}, Gsdmd (gasdermin D) [NCBI Gene 69146] {aka 1810036L03Rik, DF5L, Dfna5l, GsdmD-1, Gsdmdc1, M2-4}, Il6 (interleukin 6) [NCBI Gene 16193] {aka Il-6}, Col1a2 (collagen, type I, alpha 2) [NCBI Gene 12843] {aka Col1a-2, Cola-2, Cola2, oim}, Aif1 (allograft inflammatory factor 1) [NCBI Gene 29427] {aka BART-1, Bart1, iba1, mrf-1}, Casp8 (caspase 8) [NCBI Gene 12370] {aka CASP-8, FLICE, MACH, Mch5}, Tnc (tenascin C) [NCBI Gene 21923] {aka C130033P17Rik, Hxb, TN, TN-C, Ten, cytotactin}, Gfap (glial fibrillary acidic protein) [NCBI Gene 24387], Il1b (interleukin 1 beta) [NCBI Gene 16176] {aka IL-1beta, Il-1b}, CD63 (CD63 molecule) [NCBI Gene 967] {aka AD1, HOP-26, ME491, MLA1, OMA81H, Pltgp40}, Casp3 (caspase 3) [NCBI Gene 12367] {aka A830040C14Rik, AC-3, CASP-3, CC3, CPP-32, CPP32}, CD9 (CD9 molecule) [NCBI Gene 928] {aka BTCC-1, DRAP-27, MIC3, MRP-1, TSPAN-29, TSPAN29}, Ms6hm (minisatellite 6 hypermutable) [NCBI Gene 17653] {aka PC-1}, TNC (tenascin C) [NCBI Gene 3371] {aka 150-225, DFNA56, GMEM, GP, HXB, JI}, CD81 (CD81 molecule) [NCBI Gene 975] {aka CVID6, S5.7, TAPA1, TSPAN28}, Vegfa (vascular endothelial growth factor A) [NCBI Gene 22339] {aka L-VEGF, Vegf, Vpf}, Igfbp2 (insulin-like growth factor binding protein 2) [NCBI Gene 16008] {aka IBP-2, Igfbp-2, mIGFBP-2}, F2 (coagulation factor II, thrombin) [NCBI Gene 2147] {aka PT, RPRGL2, THPH1}, CD5L (CD5 molecule like) [NCBI Gene 922] {aka AIM, API6, CT-2, PRO229, SP-ALPHA, Spalpha}, Grap (GRB2-related adaptor protein) [NCBI Gene 71520] {aka 8430435N19Rik}, Aif1 (allograft inflammatory factor 1) [NCBI Gene 11629] {aka AIF-1, D17H6S50E, G1, Iba1}, Lcp1 (lymphocyte cytosolic protein 1) [NCBI Gene 18826] {aka D14Ertd310e, LCP-1, Pls2, pp65}, Cd5l (CD5 antigen-like) [NCBI Gene 11801] {aka 1/6, AAC-11, AIM, Api6, CT2, Pdp}, IGFBP2 (insulin like growth factor binding protein 2) [NCBI Gene 3485] {aka IBP2, IGF-BP53}, Vwf (Von Willebrand factor) [NCBI Gene 22371] {aka 6820430P06Rik, B130011O06Rik, C630030D09, F8VWF, VWD}, MPO (myeloperoxidase) [NCBI Gene 4353], Gfap (glial fibrillary acidic protein) [NCBI Gene 14580], Nlrp3 (NLR family, pyrin domain containing 3) [NCBI Gene 216799] {aka AGTAVPRL, AII/AVP, Cias1, FCAS, FCU, MWS} - **Diseases:** ROP (MESH:D012178), retinopathy (MESH:D058437), lung inflammation (MESH:D011014), OIR (MESH:D000860), neurodegeneration (MESH:D019636), injury to (MESH:D014947), Inflammation (MESH:D007249), Hyperoxia (MESH:D018496), dry AMD (MESH:D006009), diabetic neovascular disease (MESH:D003920), lung disease (MESH:D008171), vascular diseases (MESH:D014652), cancer (MESH:D009369), neurodevelopment impairment (MESH:D060825), organ damage (MESH:D000092124), optic nerve degeneration (MESH:D009410), lung and brain injury (MESH:C567034), retinal diseases (MESH:D012164), NDI (MESH:D018500), HIV infection (MESH:D015658), lung underdevelopment (MESH:C000721289), Retinal inflammation (MESH:D012173), diabetic retinopathy (MESH:D003930), vascular disorders (MESH:D002561), BPD (MESH:D001997) - **Chemicals:** oxygen (MESH:D010100), EDTA (MESH:D004492), paraformaldehyde (MESH:C003043), PBS (MESH:D007854), DAPI (MESH:C007293), AF-488 (-), AF-647 (MESH:C569686) - **Species:** Mus musculus (house mouse, species) [taxon 10090], Rattus norvegicus (brown rat, species) [taxon 10116], Homo sapiens (human, species) [taxon 9606] - **Cell lines:** Hu — Homo sapiens (Human), Finite cell line (CVCL_B0BH), C57BL/6J — Mus musculus (Mouse), Transformed cell line (CVCL_C0MW) ## Figures 7 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12940086/full.md --- Source: https://tomesphere.com/paper/PMC12940086