{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:18:04Z","timestamp":1785406684685,"version":"3.56.0"},"posted":{"date-parts":[[2026,3,21]]},"group-title":"Biophysics","reference-count":40,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T00:00:00Z","timestamp":1774051200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["R01CA228272"],"award-info":[{"award-number":["R01CA228272"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["U54 CA268084"],"award-info":[{"award-number":["U54 CA268084"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["T32AI083216"],"award-info":[{"award-number":["T32AI083216"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["K23DK144661"],"award-info":[{"award-number":["K23DK144661"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007059","name":"Northwestern University","doi-asserted-by":"crossref","award":["Starzl Scholarship"],"award-info":[{"award-number":["Starzl Scholarship"]}],"id":[{"id":"10.13039\/100007059","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,3,21]]},"abstract":"<jats:p>Understanding how chromatin folds in three dimensions remains challenging because most experimental assays capture low-dimensional projections of an underlying, highly heterogeneous polymer. Here we present an ensemble-based interpretive framework built on the previously introduced Self-Returning Excluded Volume (SR-EV) model, a minimal generator of nucleosome-resolution chromatin conformations based on stochastic return rules and excluded-volume geometry. Despite its simplicity, SR-EV reproduces key experimental signatures across scales: heterogeneous nanoscale packing domains resembling ChromEMT and ChromSTEM observations, sparse and highly variable single-configuration contact patterns analogous to single-cell chromosome conformation capture (Hi-C), and robust ensemble-level contact enrichment consistent with topologically associating domains (TADs). In this framework, Hi-C loop and TAD signatures are interpreted as ensemble-level statistical enrichments rather than invariant features of single-cell conformations. SR-EV is explicitly designed to generate large ensembles of complete three-dimensional chromatin configurations that can be projected consistently onto two-dimensional contact maps and one-dimensional genomic profiles. By introducing architectural-protein effects only through ensemble selection rather than explicit forces, SR-EV supports a separation between intrinsic polymer geometry and regulatory bias and suggests that TAD-like features can emerge as statistical enrichments rather than deterministic three-dimensional structures. Coordination number and probe-based accessibility computed directly from SR-EV provide a unified link between three-dimensional packing, two-dimensional contact maps, and one-dimensional genomic profiles. Together, these results establish SR-EV as a minimal and physically grounded reference framework for interpreting how heterogeneous chromatin ensembles give rise to multimodal experimental observables, while remaining consistent with the fact that chromatin organization is realized in individual cells.<\/jats:p>\n                <jats:sec>\n                  <jats:title>SIGNIFICANCE<\/jats:title>\n                  <jats:p>Chromatin domains, boundaries, and contact enrichments are often interpreted as fixed structural entities, even though most experimental measurements average over large and heterogeneous cell populations. The SR-EV framework shows that many of these features can be understood as emerging from minimal geometric rules combined with ensemble-level bias, without requiring explicit molecular interactions or deterministic folding mechanisms. By distinguishing single-configuration heterogeneity from ensemble-level statistical organization \u2013 including the emergence of packing domains\u2013 SR-EV supports an interpretation in which chromatin organization is realized in individual cells but must be analyzed through ensembles. This perspective clarifies the probabilistic nature of genome architecture and provides a tractable reference framework for interpreting multimodal genomic and imaging data.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.03.19.710883","type":"posted-content","created":{"date-parts":[[2026,3,22]],"date-time":"2026-03-22T04:30:11Z","timestamp":1774153811000},"source":"Crossref","is-referenced-by-count":0,"title":["One Chromatin, Many Structures: From Ensemble Contact Maps to Single-Cell 3D Organization"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8345-7724","authenticated-orcid":false,"given":"M. A.","family":"Carignano","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering, Northwestern University and Center for Physical Genomics and Engineering, Northwestern University, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1981-1818","authenticated-orcid":false,"given":"V.","family":"Backman","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Northwestern University and Center for Physical Genomics and Engineering, Northwestern University, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1402-6714","authenticated-orcid":false,"given":"M.","family":"Kr\u00f6ger","sequence":"additional","affiliation":[{"name":"Magnetism and Interface Physics & Computational Polymer Physics, Department of Materials, ETH Zurich, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9355-7681","authenticated-orcid":false,"given":"Luay M.","family":"Almassalha","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, USA"},{"name":"Center for Physical Genomics and Engineering, Northwestern University, Evanston, Illinois, USA"},{"name":"Division of Gastroenterology and Hepatology, Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8708-0335","authenticated-orcid":false,"given":"I.","family":"Szleifer","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Northwestern University Center for Physical Genomics and Engineering, Northwestern University, United States"},{"name":"Department of Chemistry, Northwestern University, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026032408300842000_2026.03.19.710883v1.1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aag0025"},{"key":"2026032408300842000_2026.03.19.710883v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-16028-2"},{"key":"2026032408300842000_2026.03.19.710883v1.3","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adq6652"},{"key":"2026032408300842000_2026.03.19.710883v1.4","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.97604"},{"key":"2026032408300842000_2026.03.19.710883v1.5","doi-asserted-by":"publisher","DOI":"10.1126\/science.1181369"},{"key":"2026032408300842000_2026.03.19.710883v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.11.021"},{"key":"2026032408300842000_2026.03.19.710883v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/nature12593"},{"key":"2026032408300842000_2026.03.19.710883v1.8","doi-asserted-by":"publisher","DOI":"10.1038\/nature21429"},{"key":"2026032408300842000_2026.03.19.710883v1.9","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks1029"},{"key":"2026032408300842000_2026.03.19.710883v1.10","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2018.008"},{"key":"2026032408300842000_2026.03.19.710883v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/nature16496"},{"key":"2026032408300842000_2026.03.19.710883v1.12","doi-asserted-by":"publisher","DOI":"10.1038\/nature20158"},{"key":"2026032408300842000_2026.03.19.710883v1.13","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1518552112"},{"key":"2026032408300842000_2026.03.19.710883v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2016.04.085"},{"key":"2026032408300842000_2026.03.19.710883v1.15","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.25776"},{"key":"2026032408300842000_2026.03.19.710883v1.16","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-20292-z"},{"key":"2026032408300842000_2026.03.19.710883v1.17","doi-asserted-by":"publisher","DOI":"10.1080\/19491034.2016.1239684"},{"key":"2026032408300842000_2026.03.19.710883v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.05.004"},{"key":"2026032408300842000_2026.03.19.710883v1.19","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adp0855"},{"key":"2026032408300842000_2026.03.19.710883v1.20","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-016-0909-0"},{"key":"2026032408300842000_2026.03.19.710883v1.21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1006617"},{"key":"2026032408300842000_2026.03.19.710883v1.22","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceb.2024.102421"},{"key":"2026032408300842000_2026.03.19.710883v1.23","doi-asserted-by":"publisher","DOI":"10.1038\/s41576-018-0060-8"},{"key":"2026032408300842000_2026.03.19.710883v1.24","doi-asserted-by":"publisher","DOI":"10.1038\/s44320-025-00107-3"},{"key":"2026032408300842000_2026.03.19.710883v1.25","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1152\/physiol.00050.2024","article-title":"A Multiscale Perspective on Chromatin Architecture through Polymer Physics","volume":"40","year":"2025","journal-title":"Physiology"},{"key":"2026032408300842000_2026.03.19.710883v1.26","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw514"},{"key":"2026032408300842000_2026.03.19.710883v1.27","doi-asserted-by":"publisher","DOI":"10.1126\/science.aau1783"},{"key":"2026032408300842000_2026.03.19.710883v1.28","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1705685114"},{"key":"2026032408300842000_2026.03.19.710883v1.29","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.5b04086"},{"key":"2026032408300842000_2026.03.19.710883v1.30","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1816424116"},{"key":"2026032408300842000_2026.03.19.710883v1.31","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1717730115"},{"key":"2026032408300842000_2026.03.19.710883v1.32","doi-asserted-by":"crossref","first-page":"013010","DOI":"10.1103\/PRXLife.1.013010","article-title":"From Effective Interactions Extracted Using Hi-C Data to Chromosome Structures in Conventional and Inverted Nuclei","volume":"1","year":"2023","journal-title":"PRX Life"},{"key":"2026032408300842000_2026.03.19.710883v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.05.009"},{"key":"2026032408300842000_2026.03.19.710883v1.34","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aay4055"},{"key":"2026032408300842000_2026.03.19.710883v1.35","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaz3418"},{"key":"2026032408300842000_2026.03.19.710883v1.36","doi-asserted-by":"publisher","DOI":"10.1038\/nature11232"},{"key":"2026032408300842000_2026.03.19.710883v1.37","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2688"},{"key":"2026032408300842000_2026.03.19.710883v1.38","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2018.015"},{"key":"2026032408300842000_2026.03.19.710883v1.39","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-09982-5"},{"key":"2026032408300842000_2026.03.19.710883v1.40","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2151-x"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.03.19.710883","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:15:38Z","timestamp":1782278138000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.03.19.710883"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,21]]},"references-count":40,"URL":"https:\/\/doi.org\/10.64898\/2026.03.19.710883","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.1016\/j.bpj.2026.05.020","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2026,3,21]]},"subtype":"preprint"}}