{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,4,12]],"date-time":"2026-04-12T11:05:58Z","timestamp":1775991958299,"version":"3.50.1"},"posted":{"date-parts":[[2025,6,15]]},"group-title":"Neuroscience","reference-count":68,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,6,15]],"date-time":"2025-06-15T00:00:00Z","timestamp":1749945600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"NIH","award":["NIH-NINDS R21NS130319"],"award-info":[{"award-number":["NIH-NINDS R21NS130319"]}]},{"name":"The Slomo and Cindy Silvian Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,6,15]]},"abstract":"<jats:title>Summary<\/jats:title>\n                <jats:p>\n                  Pruriception is crucial for defense against external stimuli but can lead to chronic pruritus, a debilitating condition affecting millions worldwide. Our understanding of the cellular and molecular mechanisms behind the sensation of itch has been hindered by the lack of functional human models. Here, we address this limitation by developing a protocol to generate induced pruriceptors (iPruriceptors) from human pluripotent stem cells (hPSCs). We compared two differentiation approaches: a direct method via forced expression of transcription factors (TFs) in hPSCs, and a 2-step process through expression of TFs in hPSC-derived neural crest cells (NCCs). The 2-step protocol proved superior in inducing a transcriptional program that closely resembles that of human pruriceptors. Our optimized protocol employs forced expression of NGN1 and ISL1 to drive differentiation from NCCs into pruriceptors, enhancing the expression of known pruritogen receptors such as IL31RA, which pairs with OSMR, and HRH1. The induction of this transcriptional program leads to functional maturation of iPruriceptors. Accordingly, iPruriceptors exhibit robust responses to itch stimuli and\n                  <jats:italic>in vivo<\/jats:italic>\n                  -like itch pharmacology such as treatment with ABT-317, a JAK1 inhibitor tool compound, similar to those targeting intensive pruritus in atopic dermatitis (AD). Importantly, iPruriceptors can be generated without viral vectors or safe-harbor gene editing, using a PiggyBac-based transfection method that simplifies scalability. Our protocol offers a robust platform for investigating itch biology, modeling chronic pruritus, and enabling high-throughput screening for therapeutic target discovery.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Highlights<\/jats:title>\n                  <jats:list list-type=\"bullet\">\n                    <jats:list-item>\n                      <jats:p>NGN1 and ISL1 forced expression in NCCs induces rapid differentiation to iPruriceptors<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>iPruriceptors share transcriptional profile of primary human pruriceptors<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>iPruriceptors have electrophysiological responses to known pruritogens<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>iPruriceptors have JAK1-dependent IL-31\/-13 responses blocked by ABT-317<\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                <\/jats:sec>","DOI":"10.1101\/2025.06.11.659208","type":"posted-content","created":{"date-parts":[[2025,6,15]],"date-time":"2025-06-15T23:25:17Z","timestamp":1750029917000},"source":"Crossref","is-referenced-by-count":0,"title":["Induction of Human Pruriceptors from Pluripotent Stem Cells via Transcription Factors"],"prefix":"10.64898","author":[{"given":"Hisato","family":"Iriki","sequence":"first","affiliation":[{"name":"Kimberly and Eric J. Waldman Department of Dermatology, Icahn School of Medicine at Mount Sinai, New York, NY, USA"},{"name":"Mark Lebwohl Center for Neuroinflammation and Sensation, Icahn School of Medicine at Mount Sinai, New York, NY, USA"},{"name":"Marc and Jennifer Lipschultz Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruiqi","family":"Hu","sequence":"additional","affiliation":[{"name":"Nash Family Department of Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA"},{"name":"Institute for Regenerative Medicine, Icahn School of Medicine at Mount Sinai, New York, NY, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xu","family":"Li","sequence":"additional","affiliation":[{"name":"Kimberly and Eric J. Waldman Department of Dermatology, Icahn School of Medicine at Mount Sinai, New York, NY, USA"},{"name":"Mark Lebwohl Center for Neuroinflammation and Sensation, Icahn School of Medicine at Mount Sinai, New York, NY, USA"},{"name":"Marc and Jennifer Lipschultz Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erdene","family":"Baljinnyam","sequence":"additional","affiliation":[{"name":"Institute for Regenerative Medicine, Icahn School of Medicine at Mount Sinai, New York, NY, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carina","family":"Habich","sequence":"additional","affiliation":[{"name":"AbbVie Deutschland GmbH & Co KG, Neuroscience Discovery, Knollstrasse, Ludwigshafen am Rhein, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ichiro","family":"Imanishi","sequence":"additional","affiliation":[{"name":"Kimberly and Eric J. Waldman Department of Dermatology, Icahn School of Medicine at Mount Sinai, New York, NY, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Loan","family":"Miller","sequence":"additional","affiliation":[{"name":"AbbVie Inc., North Chicago, IL, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kavya","family":"Chegireddy","sequence":"additional","affiliation":[{"name":"AbbVie Bioresearch Center, Worcester, MA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laraib Iqbal","family":"Malik","sequence":"additional","affiliation":[{"name":"AbbVie Bioresearch Center, Worcester, MA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Yassky","sequence":"additional","affiliation":[{"name":"Kimberly and Eric J. Waldman Department of Dermatology, Icahn School of Medicine at Mount Sinai, New York, NY, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aaron Ver","family":"Heul","sequence":"additional","affiliation":[{"name":"Division of Allergy and Immunology, John T. Milliken Department of Medicine, Washington University School of Medicine in St. Louis, 660 S. Euclid Avenue, MSC 8122-0021-03, St. Louis, MO, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kathleen M.","family":"Smith","sequence":"additional","affiliation":[{"name":"AbbVie Bioresearch Center, Worcester, MA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eric R.","family":"Goedken","sequence":"additional","affiliation":[{"name":"AbbVie Bioresearch Center, Worcester, MA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter","family":"Reinhardt","sequence":"additional","affiliation":[{"name":"AbbVie Deutschland GmbH & Co KG, Neuroscience Discovery, Knollstrasse, Ludwigshafen am Rhein, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brian S.","family":"Kim","sequence":"additional","affiliation":[{"name":"Nash Family Department of Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA"},{"name":"Kimberly and Eric J. 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