{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,11]],"date-time":"2026-11-11T06:34:34Z","timestamp":1794378874749,"version":"4.9.0"},"reference-count":31,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,6]],"date-time":"2025-09-06T00:00:00Z","timestamp":1757116800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Background: Adeno-associated viral (AAV) vectors are the leading platform for gene therapy, but common delivery routes show limited spread to distal cortical structures, hence the utility of direct, intrathalamic infusions for broader transgene distribution. In this preliminary study, we recapitulate previous studies targeting the thalamus as a conduit to achieve cortical transgene spread and showcase novel data evaluating biodistribution of a green fluorescent protein (GFP) using cryo-fluorescence tomography (CFT). For the first time in nonhuman primates (NHPs) and coupled with magnetic resonance imaging (MRI)-guidance, we demonstrated the application of CFT as a powerful tool to map out vector distribution in the NHP brain. Methods: Briefly, a single thalamic infusion was performed in African green monkeys using ClearPoint\u2019s navigational platform to deliver an AAV serotype 2 vector containing a GFP payload. Transgene biodistribution was assessed in the left and right hemispheres using CFT and histological analysis, respectively. Results: Infusions were successfully performed with sub-millimetric target accuracy and with minimal error, achieving ~86% thalamic coverage with the largest infusion volume. Histology confirmed the presence of the GFP transgene, with the strongest signal in the cerebral gray\/white matter and internal capsule, while CFT allowed for the three-dimensional detection of the transgene starting at the site of infusion and spreading to multiple cortical regions. Conclusions: These findings suggest that by combining MRI-guided technology with CFT imaging, it is feasible to map whole-brain gene biodistribution in NHPs. This proof-of-concept study bridges the gap between cellular microscopy and MRI-guidance to provide a complete picture of disease and treatment with clinical applicability.<\/jats:p>","DOI":"10.3390\/pharmaceutics17091167","type":"journal-article","created":{"date-parts":[[2025,9,8]],"date-time":"2025-09-08T08:06:32Z","timestamp":1757318792000},"page":"1167","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["A Preliminary Study on the Accuracy of MRI-Guided Thalamic Infusion of AAV2-GFP and Biodistribution Analysis Using Cryo-Fluorescence Tomography in Nonhuman Primates"],"prefix":"10.3390","volume":"17","author":[{"given":"Ernesto A.","family":"Salegio","sequence":"first","affiliation":[{"name":"ClearPoint Neuro Inc., Solana Beach, CA 92075, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reinier","family":"Espinosa","sequence":"additional","affiliation":[{"name":"Envol Biomedical, Immokalee, FL 34142, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geary R.","family":"Smith","sequence":"additional","affiliation":[{"name":"ClearPoint Neuro Inc., Solana Beach, CA 92075, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Shoshan","sequence":"additional","affiliation":[{"name":"ClearPoint Neuro Inc., Solana Beach, CA 92075, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew","family":"Silva","sequence":"additional","affiliation":[{"name":"EMIT Imaging, Inc., Natick, MA 01760, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eli","family":"White","sequence":"additional","affiliation":[{"name":"EMIT Imaging, Inc., Natick, MA 01760, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4099-5930","authenticated-orcid":false,"given":"Jacob","family":"McDonald","sequence":"additional","affiliation":[{"name":"Envol Biomedical, Immokalee, FL 34142, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1038\/s41573-019-0012-9","article-title":"Adeno-Associated Virus Vector as a Platform for Gene Therapy Delivery","volume":"18","author":"Wang","year":"2019","journal-title":"Nat. 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