{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T05:51:43Z","timestamp":1791525103028,"version":"4.3.4"},"reference-count":45,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T00:00:00Z","timestamp":1791504000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Aging Neurosci."],"abstract":"<jats:p>Chronic neuroinflammation driven by sustained microglial and astrocyte activation contributes to Alzheimer\u2019s disease progression, yet no therapeutic strategy addresses it with precision or durability in the brain. Interleukin-10 occupies an unresolved position in this problem. Constitutive interleukin-10 overexpression in amyloid precursor protein transgenic mice increased plaque burden by suppressing microglial phagocytic activity, while interleukin-10 deletion has been reported to restrict amyloidosis in one setting and to worsen tau hyperphosphorylation in another. We interpret this pattern as evidence that interleukin-10 carries opposite signs on the two principal pathologies of Alzheimer\u2019s disease, so that any therapy holding it at a fixed concentration is committed in advance to being wrong about one of them. We therefore propose a falsifiable hypothesis: conditional, inflammation-gated interleukin-10 expression may suppress damaging inflammatory signaling while limiting the impairment of microglial phagocytosis associated with continuous exposure. We present a design framework for a synthetic gene circuit combining an adeno-associated viral vector for delivery to the central nervous system, a synthetic Notch receptor that detects membrane-bound tumor necrosis factor alpha on activated microglia through juxtacrine contact, and a PiggyBac transposon intended to integrate the circuit into the astrocyte genome. Receptor engagement is expected to release a transcription factor that drives interleukin-10 from a synthetic promoter, with transcription ceasing as the stimulus is withdrawn. We provide a quantitative packaging analysis showing that the circuit requires a compacted vector design to fit within viral capacity, and we identify three unvalidated assumptions: the recognition domain does not yet exist, ligand surface density on activated microglia is unmeasured, and shed soluble tumor necrosis factor alpha is predicted to act as a competitive inhibitor of receptor activation. A computational analysis of simulated resting-state electroencephalography, in which 45 of 171 electrode pairs differed after false discovery rate correction, is presented as a fully specified analysis workflow for delta-band connectivity as a secondary outcome measure rather than as evidence of effect, and yields effect sizes indicating that a rodent study would need roughly 11 animals per group. A four-arm validation plan is specified that separates the contribution of conditional regulation from that of genomic integration.<\/jats:p>","DOI":"10.3389\/fnagi.2026.1866161","type":"journal-article","created":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T05:30:10Z","timestamp":1791523810000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["A SynNotch-PiggyBac gene circuit for inflammation-responsive IL-10 delivery in Alzheimer\u2019s disease: a hypothesis and design framework"],"prefix":"10.3389","volume":"18","author":[{"given":"Suhani","family":"Gupta","sequence":"first","affiliation":[{"name":"Neuro Health Alliance","place":["Dublin, CA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronav","family":"Gupta","sequence":"additional","affiliation":[{"name":"IGNITE Pathways","place":["Dublin, CA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,10,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1111\/j.2517-6161.1995.tb02031.x","article-title":"Controlling the false discovery rate: A practical and powerful approach to multiple testing.","volume":"57","author":"Benjamini","year":"1995","journal-title":"J. R. Stat. Soc. Series B Stat. Methodol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"eadm8563","DOI":"10.1126\/scitranslmed.adm8563","article-title":"Microglia-specific IL-10 gene delivery inhibits neuroinflammation and neurodegeneration in a mouse model of Parkinson\u2019s disease.","volume":"16","author":"Bido","year":"2024","journal-title":"Sci. Transl. Med."},{"key":"B3","doi-asserted-by":"publisher","first-page":"e086813","DOI":"10.1002\/alz.086813","article-title":"MRI-driven analysis of EEG connectivity in Alzheimer\u2019s disease and behavioral frontotemporal dementia","volume":"20","author":"Cecchetti","year":"2024","journal-title":"Alzheimers Dement"},{"key":"B4","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1016\/j.neuron.2014.11.020","article-title":"IL-10 alters immunoproteostasis in APP mice, increasing plaque burden and worsening cognitive behavior.","volume":"85","author":"Chakrabarty","year":"2015","journal-title":"Neuron"},{"key":"B5","doi-asserted-by":"publisher","first-page":"eads3972","DOI":"10.1126\/science.ads3972","article-title":"Targeting amyloid-\u03b2 pathology by chimeric antigen receptor astrocyte (CAR-A) therapy","volume":"391","author":"Chen","year":"2026","journal-title":"Science"},{"key":"B6","doi-asserted-by":"publisher","first-page":"667","DOI":"10.1038\/mt.2014.254","article-title":"Hybrid nonviral\/viral vector systems for improved piggyBac DNA transposon in vivo delivery.","volume":"23","author":"Cooney","year":"2015","journal-title":"Mol. Ther."},{"key":"B7","doi-asserted-by":"publisher","first-page":"2167","DOI":"10.1016\/S0140-6736(26)00459-9","article-title":"Efficacy and safety of oral semaglutide 14 mg (flexible dose) in early-stage symptomatic Alzheimer\u2019s disease (evoke and evoke+): two phase 3, randomised, placebo-controlled trials","volume":"407","author":"Cummings","year":"2026","journal-title":"Lancet"},{"key":"B8","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1002\/hep.27842","article-title":"Modeling correction of severe urea cycle defects in the growing murine liver using a hybrid recombinant adeno-associated virus\/piggyBac transposase gene delivery system","volume":"62","author":"Cunningham","year":"2015","journal-title":"Hepatology"},{"key":"B9","doi-asserted-by":"publisher","first-page":"1073","DOI":"10.1016\/j.cell.2018.05.003","article-title":"Disease-associated microglia: A universal immune sensor of neurodegeneration.","volume":"173","author":"Deczkowska","year":"2018","journal-title":"Cell"},{"key":"B10","doi-asserted-by":"publisher","first-page":"312","DOI":"10.1038\/s41593-020-00783-4","article-title":"Reactive astrocyte nomenclature, definitions, and future directions.","volume":"24","author":"Escartin","year":"2021","journal-title":"Nat. Neurosci."},{"key":"B11","doi-asserted-by":"publisher","first-page":"e105","DOI":"10.1016\/S2468-2667(21)00249-8","article-title":"Estimation of the global prevalence of dementia in 2019 and forecasted prevalence in 2050: an analysis for the Global Burden of Disease Study 2019","volume":"7","year":"2022","journal-title":"Lancet Public Health"},{"key":"B12","doi-asserted-by":"publisher","first-page":"198","DOI":"10.1038\/s41434-019-0075-6","article-title":"Astrocyte-selective AAV gene therapy through the endogenous GFAP promoter results in robust transduction in the rat spinal cord following injury.","volume":"26","author":"Griffin","year":"2019","journal-title":"Gene Ther."},{"key":"B13","doi-asserted-by":"publisher","first-page":"534","DOI":"10.1016\/j.neuron.2014.12.068","article-title":"Il10 deficiency rebalances innate immunity to mitigate Alzheimer-like pathology.","volume":"85","author":"Guillot-Sestier","year":"2015","journal-title":"Neuron"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/s41434-026-00617-1","article-title":"AAV2 capsid clearance and neuronal trafficking dynamics in the central nervous system.","author":"Gullapalli","year":"2026","journal-title":"Gene Ther."},{"key":"B15","doi-asserted-by":"publisher","first-page":"4864","DOI":"10.1038\/s41598-021-84210-z","article-title":"Glial cell type-specific gene expression in the mouse cerebrum using the piggyBac system and in utero electroporation.","volume":"11","author":"Hamabe-Horiike","year":"2021","journal-title":"Sci. Rep."},{"key":"B16","doi-asserted-by":"publisher","first-page":"1575453","DOI":"10.3389\/fnins.2025.1575453","article-title":"The roles of microglia and astrocytes in neuroinflammation of Alzheimer\u2019s disease.","volume":"19","author":"Han","year":"2025","journal-title":"Front. Neurosci."},{"key":"B17","doi-asserted-by":"publisher","first-page":"109530","DOI":"10.1016\/j.jneumeth.2022.109530","article-title":"Characterization of the GfaABC1D promoter to selectively target astrocytes in the rhesus macaque brain.","volume":"372","author":"Heffernan","year":"2022","journal-title":"J. Neurosci. Methods"},{"key":"B18","doi-asserted-by":"publisher","first-page":"388","DOI":"10.1016\/S1474-4422(15)70016-5","article-title":"Neuroinflammation in Alzheimer\u2019s disease.","volume":"14","author":"Heneka","year":"2015","journal-title":"Lancet Neurol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1166","DOI":"10.1126\/science.abc1855","article-title":"T cell circuits that sense antigen density with an ultrasensitive threshold.","volume":"371","author":"Hernandez-Lopez","year":"2021","journal-title":"Science"},{"key":"B20","doi-asserted-by":"publisher","first-page":"1051559","DOI":"10.3389\/fneur.2022.1051559","article-title":"Kinetics and durability of transgene expression after intrastriatal injection of AAV9 vectors.","volume":"13","author":"Hollidge","year":"2022","journal-title":"Front. Neurol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1186\/1750-1326-6-69","article-title":"Elevated CSF levels of TACE activity and soluble TNF receptors in subjects with mild cognitive impairment and patients with Alzheimer\u2019s disease.","volume":"6","author":"Jiang","year":"2011","journal-title":"Mol. Neurodegener."},{"key":"B22","doi-asserted-by":"publisher","first-page":"1276","DOI":"10.1016\/j.cell.2017.05.018","article-title":"A unique microglia type associated with restricting development of Alzheimer\u2019s disease.","volume":"169","author":"Keren-Shaul","year":"2017","journal-title":"Cell"},{"key":"B23","doi-asserted-by":"publisher","first-page":"826","DOI":"10.3390\/biomedicines14040826","article-title":"IL-10-STAT3-dependent transcriptional regulation in microglia: Alzheimer\u2019s disease and neuroinflammation.","volume":"14","author":"Kim","year":"2026","journal-title":"Biomedicines"},{"key":"B24","doi-asserted-by":"publisher","first-page":"2327","DOI":"10.1002\/alz.12610","article-title":"Role of chronic neuroinflammation in neuroplasticity and cognitive function: A hypothesis.","volume":"18","author":"Lecca","year":"2022","journal-title":"Alzheimers Dement."},{"key":"B25","doi-asserted-by":"publisher","first-page":"4343","DOI":"10.3390\/ijms26094343","article-title":"In situ synNotch-programmed astrocytes sense and attenuate neuronal apoptosis.","volume":"26","author":"Liang","year":"2025","journal-title":"Int. J. Mol. Sci."},{"key":"B26","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1186\/s12974-016-0763-8","article-title":"Balancing the immune response in the brain: IL-10 and its regulation.","volume":"13","author":"Lobo-Silva","year":"2016","journal-title":"J. Neuroinflammation"},{"key":"B27","doi-asserted-by":"publisher","first-page":"104","DOI":"10.3390\/ph18010104","article-title":"Targeting cytokine-mediated inflammation in brain disorders: Developing new treatment strategies.","volume":"18","author":"Mallick","year":"2025","journal-title":"Pharmaceuticals"},{"key":"B28","doi-asserted-by":"publisher","first-page":"19184","DOI":"10.1073\/pnas.1216473109","article-title":"Hyperactive self-inactivating piggyBac for transposase-enhanced pronuclear microinjection transgenesis.","volume":"109","author":"Marh","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"B29","doi-asserted-by":"publisher","first-page":"639","DOI":"10.1016\/j.neurobiolaging.2006.03.013","article-title":"NSAIDs and Alzheimer disease: Epidemiological, animal model and clinical studies.","volume":"28","author":"McGeer","year":"2007","journal-title":"Neurobiol. Aging"},{"key":"B30","doi-asserted-by":"publisher","first-page":"780","DOI":"10.1016\/j.cell.2016.01.012","article-title":"Engineering customized cell sensing and response behaviors using synthetic notch receptors.","volume":"164","author":"Morsut","year":"2016","journal-title":"Cell"},{"key":"B31","doi-asserted-by":"publisher","first-page":"1708","DOI":"10.1038\/s41591-026-04273-1","article-title":"The TREM2 agonistic antibody AL002 in early Alzheimer\u2019s disease: A phase 2 randomized trial.","volume":"32","author":"Mummery","year":"2026","journal-title":"Nat. Med."},{"key":"B32","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1016\/S0896-6273(03)00434-3","article-title":"Triple-transgenic model of Alzheimer\u2019s disease with plaques and tangles: Intracellular A\u03b2 and synaptic dysfunction.","volume":"39","author":"Oddo","year":"2003","journal-title":"Neuron"},{"key":"B33","doi-asserted-by":"publisher","first-page":"1496235","DOI":"10.3389\/fnagi.2025.1496235","article-title":"Functional and effective EEG connectivity patterns in Alzheimer\u2019s disease and mild cognitive impairment: A systematic review.","volume":"17","author":"Paitel","year":"2025","journal-title":"Front. Aging Neurosci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"1017","DOI":"10.3390\/biom10071017","article-title":"The regulatory role of IL-10 in neurodegenerative diseases.","volume":"10","author":"Porro","year":"2020","journal-title":"Biomolecules"},{"key":"B35","doi-asserted-by":"publisher","first-page":"eadl4237","DOI":"10.1126\/science.adl4237","article-title":"Programming tissue-sensing T cells that deliver therapies to the brain.","volume":"386","author":"Simic","year":"2024","journal-title":"Science"},{"key":"B36","doi-asserted-by":"publisher","first-page":"1287","DOI":"10.1038\/s41587-022-01638-y","article-title":"Tension-tuned receptors for synthetic mechanotransduction and intercellular force detection.","volume":"41","author":"Sloas","year":"2023","journal-title":"Nat. Biotechnol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.omtn.2017.03.009","article-title":"A microRNA124 target sequence restores astrocyte specificity of gfaABC1D-driven transgene expression in AAV-mediated gene transfer.","volume":"8","author":"Taschenberger","year":"2017","journal-title":"Mol. Ther. Nucleic. Acids"},{"key":"B38","doi-asserted-by":"publisher","first-page":"e101305","DOI":"10.1002\/alz70859_101305","article-title":"A phase I clinical trial of AAV2-BDNF gene therapy for Alzheimer\u2019s disease: Findings","volume":"21","author":"Tuszynski","year":"2025","journal-title":"Alzheimers Dement"},{"key":"B39","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1056\/NEJMoa2212948","article-title":"Lecanemab in early Alzheimer\u2019s disease.","volume":"388","author":"van Dyck","year":"2023","journal-title":"N. Engl. J. Med."},{"key":"B40","doi-asserted-by":"publisher","first-page":"698002","DOI":"10.3389\/fcell.2021.698002","article-title":"Non-codon optimized piggyBac transposase induces developmental brain aberrations: A call for in vivo analysis.","volume":"9","author":"Vierl","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"161","DOI":"10.1186\/s12974-021-02211-1","article-title":"Interleukin-10 deficiency exacerbates inflammation-induced tau pathology.","volume":"18","author":"Weston","year":"2021","journal-title":"J. Neuroinflammation"},{"key":"B42","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1038\/s42003-020-0848-x","article-title":"Engineering of an enhanced synthetic Notch receptor by reducing ligand-independent activation.","volume":"3","author":"Yang","year":"2020","journal-title":"Commun. Biol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"115363","DOI":"10.1016\/j.addr.2024.115363","article-title":"Adeno-associated virus vector delivery to the brain: Technology advancements and clinical applications.","volume":"211","author":"Ye","year":"2024","journal-title":"Adv. Drug Deliv. Rev."},{"key":"B44","doi-asserted-by":"publisher","first-page":"1531","DOI":"10.1073\/pnas.1008322108","article-title":"A hyperactive piggyBac transposase for mammalian applications.","volume":"108","author":"Yusa","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"B45","doi-asserted-by":"publisher","first-page":"1431","DOI":"10.1016\/j.cell.2022.03.023","article-title":"Modular design of synthetic receptors for programmed gene regulation in cell therapies.","volume":"185","author":"Zhu","year":"2022","journal-title":"Cell"}],"container-title":["Frontiers in Aging Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnagi.2026.1866161\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T05:30:11Z","timestamp":1791523811000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnagi.2026.1866161\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,9]]},"references-count":45,"alternative-id":["10.3389\/fnagi.2026.1866161"],"URL":"https:\/\/doi.org\/10.3389\/fnagi.2026.1866161","relation":{},"ISSN":["1663-4365"],"issn-type":[{"value":"1663-4365","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,9]]},"article-number":"1866161"}}