{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:28:19Z","timestamp":1785403699715,"version":"3.56.0"},"reference-count":26,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T00:00:00Z","timestamp":1769472000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T00:00:00Z","timestamp":1769472000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000897","name":"Cystic Fibrosis Foundation","doi-asserted-by":"publisher","award":["CFF-WHITELE20A0"],"award-info":[{"award-number":["CFF-WHITELE20A0"]}],"id":[{"id":"10.13039\/100000897","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mSphere"],"published-print":{"date-parts":[[2026,1,27]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Intrastrain genetic and phenotypic heterogeneity of\n                      <jats:italic toggle=\"yes\">Pseudomonas aeruginosa<\/jats:italic>\n                      is a hallmark of chronic lung infections in individuals with cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD). Although the coexistence of multiple\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      lineages within a single host is well documented, the impact of this heterogeneity on infection microbiogeography remains poorly understood. We previously showed that loss of the lipopolysaccharide (LPS) O-specific antigen (OSA) alters\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      aggregate assembly. Since OSA-deficient variants are common in chronic pulmonary infections and associated with increased pathogenesis and immune evasion, we investigated whether intrastrain OSA diversity shapes infection microbiogeography. We constructed mixed populations containing equal ratios of OSA-deficient variants and wild-type (WT) cells and examined aggregate assembly and population structures in a synthetic CF sputum model (SCFM2). To assess OSA heterogeneity\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      , we used a murine pneumonia model combined with hybridization chain reaction (HCR) RNA-FISH and whole-tissue clearing to visualize spatial organization in the airways. In SCFM2, OSA-deficient variants increased total population size, reduced WT aggregate size, and altered spatial organization. We employed 2-plex HCR RNA-FISH to distinguish WT and OSA-deficient variants in murine lungs. Interestingly, in contrast to\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      conditions, OSA-deficient cells led to significantly larger WT aggregates in the airways. These findings highlight the role of intrastrain genetic heterogeneity in shaping infection microbiogeography and provide a framework for understanding how population dynamics influence microbial physiology and host-pathogen interactions at the micron scale.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Intrastrain genetic and phenotypic diversity within\n                        <jats:italic toggle=\"yes\">Pseudomonas aeruginosa<\/jats:italic>\n                        populations is common in chronic pulmonary infections. While this intrastrain heterogeneity is a hallmark of chronic infection, its consequences for the spatial organization of\n                        <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                        within the airways remain unclear. Here, we demonstrate that the loss of O-specific antigen in a subpopulation of\n                        <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                        significantly alters the spatial architecture of\n                        <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                        , without changing the total population size or composition. Using a combination of tissue clearing and hybridization chain reaction RNA-FISH in a murine lung infection model, we mapped the localization of genetically distinct\n                        <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                        variants in mixed populations\n                        <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                        . These findings reveal that genetic diversification within a strain can reshape the infection landscape at the micron scale, highlighting the overlooked role of intrastrain dynamics in shaping the microbiogeography of infections and influencing host-pathogen interactions.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/msphere.00657-25","type":"journal-article","created":{"date-parts":[[2025,12,16]],"date-time":"2025-12-16T14:00:29Z","timestamp":1765893629000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Whole-tissue imaging reveals intrastrain diversity shapes the spatial organization of\n                    <i>Pseudomonas aeruginosa<\/i>\n                    in a murine infection model"],"prefix":"10.1128","volume":"11","author":[{"given":"H. L.","family":"Fraser","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03qt6ba18","id-type":"ROR","asserted-by":"publisher"}],"name":"Biology Department, College of Arts and Sciences, Georgia State University","place":["Atlanta, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9616-5947","authenticated-orcid":true,"given":"D. A.","family":"Moustafa","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02gars961","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Pediatrics, Division of Pulmonary, Asthma, Cystic Fibrosis, and Sleep, Emory University School of Medicine","place":["Atlanta, USA"]},{"name":"Emory+Children's Cystic Fibrosis Center","place":["Atlanta, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5285-5188","authenticated-orcid":true,"given":"J. B.","family":"Goldberg","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02gars961","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Pediatrics, Division of Pulmonary, Asthma, Cystic Fibrosis, and Sleep, Emory University School of Medicine","place":["Atlanta, USA"]},{"name":"Emory+Children's Cystic Fibrosis Center","place":["Atlanta, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9358-6087","authenticated-orcid":false,"given":"S.","family":"Azimi","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03qt6ba18","id-type":"ROR","asserted-by":"publisher"}],"name":"Biology Department, College of Arts and Sciences, Georgia State University","place":["Atlanta, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-090817-062503"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1136\/thoraxjnl-2021-217576"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-022-00683-3"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-020-00477-5"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmi.2019.06.011"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcf.2015.03.003"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1136\/thoraxjnl-2017-210134"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcf.2010.03.001"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1136\/thx.2005.045146"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejrad.2022.110421"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1002\/ppul.21011"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41396-020-0652-0"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1098\/rspb.2020.2272"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1419677112"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00860-21"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0008439"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcf.2019.04.009"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02914-18"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.00055-25"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1814023115"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2406234121"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1111\/mpp.12864"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2221542120"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1242\/dev.140137"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00796-16"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2022.101968"}],"container-title":["mSphere"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/msphere.00657-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/msphere.00657-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T14:01:13Z","timestamp":1769522473000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/msphere.00657-25"}},"subtitle":[],"editor":[{"given":"James P.","family":"O'Gara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,1,27]]},"references-count":26,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2026,1,27]]}},"alternative-id":["10.1128\/msphere.00657-25"],"URL":"https:\/\/doi.org\/10.1128\/msphere.00657-25","relation":{},"ISSN":["2379-5042"],"issn-type":[{"value":"2379-5042","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,27]]},"assertion":[{"value":"2025-10-27","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-13","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-16","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00657-25"}}