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Yet the structural principles that organize these communities and their relationship to drug tolerance remain poorly understood\n                  <jats:sup>2<\/jats:sup>\n                  , including in major human pathogens such as\n                  <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                  (\n                  <jats:italic>Mtb<\/jats:italic>\n                  )\n                  <jats:sup>3,4<\/jats:sup>\n                  . Here we show that submerged\n                  <jats:italic>Mtb<\/jats:italic>\n                  biofilms form organized multicellular architectures in which bacterial cords are integrated with a composite extracellular matrix. The virulence lipid phthiocerol dimycocerosate (PDIM) governs the higher-order organization of cords, proteinaceous matrix interactions support cohesion and attachment, an amyloid-like component contributes to biofilm integrity and establishment, and the ESX-1 secretion system increases matrix biochemical complexity. Biofilm growth conferred antibiotic tolerance across genetic backgrounds, while PDIM-dependent organization provided additional protection against first-line anti-tuberculosis drugs, most clearly isoniazid. Targeting amyloid assembly with epigallocatechin gallate impaired biofilm establishment without inhibiting planktonic growth. Together, these findings establish a cording-centred framework linking bacterial surface composition, cellular organization and extracellular matrix assembly to antibiotic tolerance, and show how resolving higher-order biofilm architecture can reveal processes amenable to chemical perturbation.\n                <\/jats:p>","DOI":"10.1101\/2025.11.07.687260","type":"posted-content","created":{"date-parts":[[2025,11,8]],"date-time":"2025-11-08T04:45:14Z","timestamp":1762577114000},"source":"Crossref","is-referenced-by-count":2,"title":["Higher-order organization of bacterial cords and an amyloid-like matrix define the\n                  <i>Mycobacterium tuberculosis<\/i>\n                  biofilm"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1104-9486","authenticated-orcid":false,"given":"Bei Shi","family":"Lee","sequence":"first","affiliation":[{"name":"Epidemiology, Biostatistics and Prevention Institute, University of Zurich, Zurich, 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