{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T21:17:08Z","timestamp":1768511828206,"version":"3.49.0"},"posted":{"date-parts":[[2024,5,10]]},"group-title":"Microbiology","reference-count":47,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,5,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Treatment of\n                  <jats:italic>Mycobacterium abscessus<\/jats:italic>\n                  pulmonary disease requires multiple antibiotics including intravenous \u03b2-lactams (e.g., imipenem, meropenem).\n                  <jats:italic>M. abscessus<\/jats:italic>\n                  produces a \u03b2-lactamase (Bla\n                  <jats:sub>Mab<\/jats:sub>\n                  ) that inactivates \u03b2-lactam drugs but less efficiently carbapenems. Due to intrinsic and acquired resistance in\n                  <jats:italic>M. abscessus<\/jats:italic>\n                  and poor clinical outcomes, it is critical to understand the development of antibiotic resistance both within the host and in the setting of outbreaks.\n                <\/jats:p>\n                <jats:p>\n                  We compared serial longitudinally collected\n                  <jats:italic>M. abscessus<\/jats:italic>\n                  subsp.\n                  <jats:italic>massiliense<\/jats:italic>\n                  isolates from the index case of a CF center outbreak and four outbreak-related strains. We found strikingly high imipenem resistance in the later patient isolates, including the outbreak strain (MIC &gt;512 \u00b5g\/ml). The phenomenon was recapitulated upon exposure of intracellular bacteria to imipenem. Addition of the \u03b2-lactamase inhibitor avibactam abrogated the resistant phenotype. Imipenem resistance was caused by an increase in \u03b2-lactamase activity and increased\n                  <jats:italic>bla<\/jats:italic>\n                  <jats:sub>Mab<\/jats:sub>\n                  mRNA level. Concurrent increase in transcription of preceding\n                  <jats:italic>ppiA<\/jats:italic>\n                  gene indicated upregulation of the entire operon in the resistant strains.\n                <\/jats:p>\n                <jats:p>\n                  Deletion of the porin\n                  <jats:italic>mspA<\/jats:italic>\n                  coincided with the first increase in MIC (from 8 to 32 \u00b5g\/ml). A frameshift mutation in\n                  <jats:italic>msp2<\/jats:italic>\n                  responsible for the rough colony morphology, and a SNP in ATP-dependent helicase\n                  <jats:italic>hrpA<\/jats:italic>\n                  co-occurred with the second increase in MIC (from 32 to 256 \u00b5g\/ml). Increased Bla\n                  <jats:sub>Mab<\/jats:sub>\n                  expression and enzymatic activity may have been due to altered regulation of the\n                  <jats:italic>ppiA<\/jats:italic>\n                  -\n                  <jats:italic>bla<\/jats:italic>\n                  <jats:sub>Mab<\/jats:sub>\n                  operon by the mutated HrpA alone, or in combination with other genes described above. This work supports using carbapenem\/\u03b2-lactamase inhibitor combinations for treating\n                  <jats:italic>M. abscessus<\/jats:italic>\n                  , particularly imipenem resistant strains.\n                <\/jats:p>","DOI":"10.1101\/2024.05.08.593223","type":"posted-content","created":{"date-parts":[[2024,5,10]],"date-time":"2024-05-10T10:20:13Z","timestamp":1715336413000},"source":"Crossref","is-referenced-by-count":2,"title":["Evolution towards extremely high \u03b2-lactam resistance in\n                  <i>Mycobacterium abscessus<\/i>\n                  outbreak strains"],"prefix":"10.64898","author":[{"given":"Eva","family":"le Run","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0615-3257","authenticated-orcid":false,"given":"Herv\u00e9","family":"Tettelin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3207-5464","authenticated-orcid":false,"given":"Steven M.","family":"Holland","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2847-7428","authenticated-orcid":false,"given":"Adrian M.","family":"Zelazny","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024051404351050000_2024.05.08.593223v1.1","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.00535-2020"},{"key":"2024051404351050000_2024.05.08.593223v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccm.2014.10.002"},{"key":"2024051404351050000_2024.05.08.593223v1.3","doi-asserted-by":"crossref","unstructured":"Ratnatunga CN , Lutzky VP , Kupz A , Doolan DL , Reid DW , Field M , et al. 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