{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:34:46Z","timestamp":1785418486302,"version":"3.56.0"},"posted":{"date-parts":[[2026,4,6]]},"group-title":"Pharmacology and Toxicology","reference-count":51,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T00:00:00Z","timestamp":1775433600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"award":["R01HD113193-02"],"award-info":[{"award-number":["R01HD113193-02"]}],"id":[{"id":"https:\/\/ror.org\/04byxyr05","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,4,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>P-glycoprotein (P-gp\/ABCB1) is a key efflux transporter that maintains barrier integrity by clearing xenobiotics and toxic metabolites. At the feto-maternal interface, trophoblast-derived extracellular vesicles (CTC-EVs) naturally and transiently transfer functional P-gp to maternal decidual cells, restoring lost and or reduced P-gp function (exofection) to sustain pregnancy homeostasis. A similar loss of P-gp at the blood brain barrier (BBB) contributes to impaired amyloid-\u03b2 (A\u03b2) clearance and neuroinflammation in Alzheimer\u2019s disease. We investigated whether CTC-EV-mediated exofection could restore P-gp function in human brain endothelial cells (hBECs) and enhance A\u03b2 clearance under inflammatory and neurodegenerative conditions.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>CTC-EVs were isolated and characterized by nanoparticle tracking analysis and western blotting for P-gp and EV markers. Transcriptomic profiling of CTC-EVs identified enrichment of transporter-related genes, including solute carriers and ABC transporters, along with inflammatory mediators. Network analysis revealed coordinated modules linking EV cargo to transporter regulation, endocytosis\/trafficking pathways, and inflammatory remodeling processes converging on BBB efflux activity. hBECs were exposed to LPS (500 ng\/mL, 48 h) with or without CTC-EVs. P-gp expression was assessed by immunofluorescence (mean fluorescence intensity, MFI) and western blotting, while functional efflux was measured using Calcein-AM assays. A\u03b2 oligomer transport was evaluated using a transwell hBEC model. In vivo, 3xTg-AD mice received intravenous CTC-EVs (1\u00d710L\/day for 5 days), followed by assessment of P-gp expression, A\u03b2 burden, and neuroinflammatory markers. Pharmacokinetic studies in P-gp knockout mice were conducted to confirm functional transporter recovery.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>LPS exposure significantly reduced P-gp expression in hBECs (41.3% decrease in MFI, p=0.0084), which was restored by CTC-EVs (46.7% increase vs. LPS, p=0.0121). Exofection increased P-gp by a 2.1-fold following EV treatment as determined by western blot. Functional assays demonstrated enhanced efflux, with a 38.5% reduction in intracellular Calcein fluorescence (p&lt;0.001). Network-informed mechanisms supported coordinated regulation of transporter and trafficking pathways. CTC-EVs improved A\u03b2 transport across inflamed hBEC monolayers. In vivo, EV-treated 3xTg-AD mice exhibited increased P-gp expression in the frontal cortex (38.6%) and hippocampus (42.1%), reduced A\u03b2 plaque burden (27.9%), and decreased inflammatory markers (IL-1\u03b2 and TNF-\u03b1, p&lt;0.05). In P-gp knockout mice, EVs reduced brain drug accumulation by 22.4% (p=0.032), confirming restoration of transporter function.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>CTC derived EVs are natural carriers of functional transporter proteins and restore efflux capacity in compromised endothelial barriers. Integration of transcriptomic and network analyses highlights coordinated regulation of transporter, trafficking, and inflammatory pathways underlying exofection. This reproductive biology inspired strategy offers a promising therapeutic approach for enhancing A\u03b2 clearance and mitigating neuroinflammation in Alzheimer\u2019s disease.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.04.02.716001","type":"posted-content","created":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T08:45:12Z","timestamp":1775465112000},"source":"Crossref","is-referenced-by-count":0,"title":["Exofection as a Therapeutic Modality: Restoring P-gp Activity via Trophoblast-Derived EV in Neuroinflammatory Disorders"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5300-2083","authenticated-orcid":false,"given":"Ananth K","family":"Kammala","sequence":"first","affiliation":[{"name":"Division of Basic Science and Translational Research, Department of Obstetrics & Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madhuri","family":"Tatiparthy","sequence":"additional","affiliation":[{"name":"Division of Basic Science and Translational Research, Department of Obstetrics & Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sravan Gopalkrishna Shetty","family":"Sreenivasmurthy","sequence":"additional","affiliation":[{"name":"Mitchell Center for Neurodegenerative Diseases, Department of Neurology, 301 University Blvd., The University of Texas Medical Branch at Galveston, Galveston, TX, 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Klarissa H","family":"Garza","sequence":"additional","affiliation":[{"name":"Mitchell Center for Neurodegenerative Diseases, Department of Neurology, 301 University Blvd., The University of Texas Medical Branch at Galveston, Galveston, TX, 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaneilahi","family":"Budhwani","sequence":"additional","affiliation":[{"name":"Mitchell Center for Neurodegenerative Diseases, Department of Neurology, 301 University Blvd., The University of Texas Medical Branch at Galveston, Galveston, TX, 77555, USA"},{"name":"Department of Neurology, University of Texas Medical Branch, Galveston, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lauren","family":"Richardson","sequence":"additional","affiliation":[{"name":"Division of Basic Science and Translational Research, Department of Obstetrics & Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramkumar","family":"Menon","sequence":"additional","affiliation":[{"name":"Division of Basic Science and Translational Research, Department of Obstetrics & Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Balaji","family":"Krishnan","sequence":"additional","affiliation":[{"name":"Mitchell Center for Neurodegenerative Diseases, Department of Neurology, 301 University Blvd., The University of Texas Medical Branch at Galveston, Galveston, TX, 77555, USA"},{"name":"Department of Neurology, University of Texas Medical Branch, Galveston, TX, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026040804351274000_2026.04.02.716001v1.1","first-page":"5","article-title":"Vol","volume":"21","author":"Association","year":"2025","journal-title":"As. 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