{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T00:22:30Z","timestamp":1775175750753,"version":"3.50.1"},"reference-count":36,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,1,13]],"date-time":"2023-01-13T00:00:00Z","timestamp":1673568000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"MUR PNRR","award":["PE00000007"],"award-info":[{"award-number":["PE00000007"]}]},{"name":"MUR PNRR","award":["GR-2018-12367178"],"award-info":[{"award-number":["GR-2018-12367178"]}]},{"name":"RICERCA FINALIZZATA","award":["PE00000007"],"award-info":[{"award-number":["PE00000007"]}]},{"name":"RICERCA FINALIZZATA","award":["GR-2018-12367178"],"award-info":[{"award-number":["GR-2018-12367178"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Even though Everolimus has been investigated in a phase II randomized trial as a host-directed therapy (HDT) to treat tuberculosis (TB), an oncological patient treated with Everolimus for a neuroendocrine pancreatic neoplasia developed active TB twice and a non-tuberculous mycobacterial (NTM) infection in a year and a half time span. To investigate this interesting case, we isolated and genotypically characterized the Mycobacterium tuberculosis (Mtb) clinical strain from the patient and tested the effect of Everolimus on its viability in an axenic culture and in a peripheral blood mononuclear cell (PBMCs) infection model. To exclude strain-specific resistance, we tested the activity of Everolimus against Mtb strains of ancient and modern lineages. Furthermore, we investigated the Everolimus effect on ROS production and autophagy modulation during Mtb infection. Everolimus did not have a direct effect on mycobacteria viability and a negligible effect during Mtb infection in host cells, although it stimulated autophagy and ROS production. Despite being a biologically plausible HDT against TB, Everolimus does not exert a direct or indirect activity on Mtb. This case underlines the need for a careful approach to drug repurposing and implementation and the importance of pre-clinical experimental studies.<\/jats:p>","DOI":"10.3390\/antibiotics12010171","type":"journal-article","created":{"date-parts":[[2023,1,16]],"date-time":"2023-01-16T02:29:55Z","timestamp":1673836195000},"page":"171","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Evaluation of Everolimus Activity against Mycobacterium tuberculosis Using In Vitro Models of Infection"],"prefix":"10.3390","volume":"12","author":[{"given":"Delia Mercedes","family":"Bianco","sequence":"first","affiliation":[{"name":"Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie\u2014Sezione di Microbiologia, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4744-9820","authenticated-orcid":false,"given":"Flavio","family":"De Maio","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze di Laboratorio e Infettivologiche, Fondazione Policlinico Universitario \u201cA. Gemelli\u201d, IRCCS, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9340-3842","authenticated-orcid":false,"given":"Giulia","family":"Santarelli","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie\u2014Sezione di Microbiologia, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ivana","family":"Palucci","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie\u2014Sezione di Microbiologia, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"},{"name":"Dipartimento di Scienze di Laboratorio e Infettivologiche, Fondazione Policlinico Universitario \u201cA. Gemelli\u201d, IRCCS, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandro","family":"Salustri","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie\u2014Sezione di Microbiologia, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1257-2295","authenticated-orcid":false,"given":"Giada","family":"Bianchetti","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Biophysics Section, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2154-319X","authenticated-orcid":false,"given":"Giuseppe","family":"Maulucci","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Biophysics Section, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Franco","family":"Citterio","sequence":"additional","affiliation":[{"name":"Renal Transplant Unit, Fondazione Policlinico Universitario A. Gemelli, IRCCS, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9780-7059","authenticated-orcid":false,"given":"Maurizio","family":"Sanguinetti","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie\u2014Sezione di Microbiologia, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"},{"name":"Dipartimento di Scienze di Laboratorio e Infettivologiche, Fondazione Policlinico Universitario \u201cA. Gemelli\u201d, IRCCS, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Enrica","family":"Tamburrini","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze di Laboratorio e Infettivologiche, Fondazione Policlinico Universitario \u201cA. Gemelli\u201d, IRCCS, 00168 Rome, Italy"},{"name":"Dipartimento di Sicurezza e Bioetica, Sez. Malattie Infettive, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michela","family":"Sali","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie\u2014Sezione di Microbiologia, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"},{"name":"Dipartimento di Scienze di Laboratorio e Infettivologiche, Fondazione Policlinico Universitario \u201cA. Gemelli\u201d, IRCCS, 00168 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giovanni","family":"Delogu","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie\u2014Sezione di Microbiologia, Universit\u00e0 Cattolica del Sacro Cuore, 00168 Rome, Italy"},{"name":"Mater Olbia Hospital, 07026 Olbia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,13]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2022). Global Tuberculosis Report 2022."},{"key":"ref_2","unstructured":"Timothy, M., and Sterling, R. (2022, July 05). Treatment of Drug-Susceptible Pulmonary Tuberculosis in Nonpregnant Adults without HIV Infection. Available online: https:\/\/www.uptodate.com\/contents\/treatment-of-drug-susceptible-pulmonary-tuberculosis-in-nonpregnant-adults-without-hiv-infection?source=mostViewed_widget#H42."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Alipanah, N., Jarlsberg, L., Miller, C., Linh, N.N., Falzon, D., Jaramillo, E., and Nahid, P. (2018). Adherence interventions and outcomes of tuberculosis treatment: A systematic review and meta-analysis of trials and observational studies. PLoS Med., 15.","DOI":"10.1371\/journal.pmed.1002595"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Wang, M.G., Wu, S.Q., and He, J.Q. (2021). Efficacy of bedaquiline in the treatment of drug-resistant tuberculosis: A systematic review and meta-analysis. BMC Infect. Dis., 21.","DOI":"10.1186\/s12879-021-06666-8"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.cmi.2021.08.007","article-title":"Pretomanid for tuberculosis: A systematic review","volume":"28","author":"Gils","year":"2022","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1038\/nrd.2018.168","article-title":"Drug repurposing: Progress, challenges and recommendations","volume":"18","author":"Pushpakom","year":"2019","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Battah, B., Chemi, G., Butini, S., Campiani, G., Brogi, S., Delogu, G., and Gemma, S. (2019). A repurposing approach for uncovering the anti-tubercular activity of FDA-approved drugs with Potential Multi-Targeting Profiles. Molecules, 24.","DOI":"10.3390\/molecules24234373"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1038\/nrd.2017.162","article-title":"Host-directed therapies for bacterial and viral infections","volume":"17","author":"Kaufmann","year":"2018","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3042","DOI":"10.3389\/fimmu.2019.03042","article-title":"Inhibition of Transglutaminase 2 as a Potential Host-Directed Therapy Against Mycobacterium tuberculosis","volume":"10","author":"Palucci","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1016\/j.isci.2019.11.001","article-title":"Cytoplasmic RNA Sensor Pathways and Nitazoxanide Broadly Inhibit Intracellular Mycobacterium tuberculosis Growth","volume":"22","author":"Ranjbar","year":"2019","journal-title":"iScience"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Martens, U.M. (2018). Everolimus. Small Molecules in Oncology, Springer International Publishing.","DOI":"10.1007\/978-3-319-91442-8"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Ashley, D., Hernandez, J., Cao, R., To, K., Yegiazaryan, A., Abrahem, R., Nguyen, T., Owens, J., Lambros, M., and Subbian, S. (2020). Antimycobacterial Effects of Everolimus in a Human Granuloma Model. J. Clin. Med., 9.","DOI":"10.3390\/jcm9072043"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"70","DOI":"10.3389\/fmicb.2018.00070","article-title":"Harnessing the mTOR Pathway for Tuberculosis Treatment","volume":"9","author":"Singh","year":"2018","journal-title":"Front. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3018","DOI":"10.1111\/ajt.15480","article-title":"Two-year outcomes in de novo renal transplant recipients receiving everolimus-facilitated calcineurin inhibitor reduction regimen from the TRANSFORM study","volume":"19","author":"Berger","year":"2019","journal-title":"Am. J. Transplant."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Cerni, S., Shafer, D., To, K., and Venketaraman, V. (2019). Investigating the Role of Everolimus in mTOR Inhibition and Autophagy Promotion as a Potential Host-Directed Therapeutic Target in Mycobacterium tuberculosis Infection. J. Clin. Med., 8.","DOI":"10.3390\/jcm8020232"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1016\/S2213-2600(20)30448-3","article-title":"Adjunctive host-directed therapies for pulmonary tuberculosis: A prospective, open-label, phase 2, randomised controlled trial","volume":"9","author":"Wallis","year":"2021","journal-title":"Lancet Respir. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"107","DOI":"10.12659\/AOT.882226","article-title":"Pulmonary embolism and reactivation of tuberculosis during everolimus therapy in a kidney transplant recipient","volume":"16","author":"Nowicki","year":"2011","journal-title":"Ann. Transplant."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"365","DOI":"10.3904\/kjim.2015.121","article-title":"Everolimus-induced activation of latent Mycobacterium tuberculosis infection in a patient with metastatic renal cell carcinoma","volume":"32","author":"Jeon","year":"2017","journal-title":"Korean J. Intern. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1905","DOI":"10.1080\/15548627.2015.1084455","article-title":"Quantitative analysis of autophagic flux by confocal pH-imaging of autophagic intermediates","volume":"11","author":"Maulucci","year":"2015","journal-title":"Autophagy"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1038\/s41419-018-0640-8","article-title":"Clinical isolates of the modern Mycobacterium tuberculosis lineage 4 evade host defense in human macrophages through eluding IL-1\u03b2-induced autophagy article","volume":"9","author":"Romagnoli","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2180","DOI":"10.1111\/ajt.13235","article-title":"Incidence and Risk Factors for Tuberculosis After Liver Transplantation in an Endemic Area: A Nationwide Population-Based Matched Cohort Study","volume":"15","author":"Chen","year":"2015","journal-title":"Am. J. Transplant."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1111\/tid.12555","article-title":"Risk of tuberculosis after lung transplantation: The value of pretransplant chest computed tomography and the impact of mTOR inhibitors and azathioprine use","volume":"18","author":"Santos","year":"2016","journal-title":"Transpl. Infect. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.phrs.2017.04.029","article-title":"Towards therapeutic drug monitoring of everolimus in cancer? Results of an exploratory study of exposure-effect relationship","volume":"121","author":"Deppenweiler","year":"2017","journal-title":"Pharmacol. Res."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Kikut, J., Jasi\u0144ska, A., Pob\u0142ocki, J., Brodowski, J., and Ma\u0142gorzata, S. (2020). Assessment and State of Nutrition of Patients with Gastroenteropancreatic Neuroendocrine Neoplasms. Nutrients, 12.","DOI":"10.3390\/nu12071961"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2634","DOI":"10.1200\/JCO.2013.49.1225","article-title":"Reactivation versus clearance of Mycobacterium tuberculosis using mammalian target of rapamycin inhibitors in patients with cancer can be context dependent","volume":"31","author":"Vale","year":"2013","journal-title":"J. Clin. Oncol. Off. J. Am. Soc. Clin. Oncol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1260","DOI":"10.1093\/infdis\/jiw347","article-title":"Human ULK1 Variation and Susceptibility to Mycobacterium tuberculosis Infection","volume":"214","author":"Horne","year":"2016","journal-title":"J. Infect. Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1016\/j.ajhg.2021.02.009","article-title":"Human ancient DNA analyses reveal the high burden of tuberculosis in Europeans over the last 2000 years","volume":"108","author":"Kerner","year":"2021","journal-title":"Am. J. Hum. Genet."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1007\/s00251-017-1017-3","article-title":"A genomic perspective on HLA evolution","volume":"70","author":"Meyer","year":"2018","journal-title":"Immunogenetics"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1504","DOI":"10.1038\/s41588-021-00935-7","article-title":"A high-resolution HLA reference panel capturing global population diversity enables multi-ancestry fine-mapping in HIV host response","volume":"53","author":"Luo","year":"2021","journal-title":"Nat. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1535","DOI":"10.1038\/ng.3704","article-title":"Mycobacterium tuberculosis lineage 4 comprises globally distributed and geographically restricted sublineages","volume":"48","author":"Stucki","year":"2016","journal-title":"Nat. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"151506","DOI":"10.1016\/j.ijmm.2021.151506","article-title":"First description of the katG gene deletion in a Mycobacterium tuberculosis clinical isolate and its impact on the mycobacterial fitness","volume":"311","author":"Cingolani","year":"2021","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1128\/JCM.01904-15","article-title":"Multicenter Evaluation of Anyplex Plus MTB\/NTM MDR-TB Assay for Rapid Detection of Mycobacterium tuberculosis Complex and Multidrug-Resistant Isolates in Pulmonary and Extrapulmonary Specimens","volume":"54","author":"Sali","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1128\/jcm.31.2.406-409.1993","article-title":"Strain Identification of Mycobacterium tuberculosis by DNA Fingerprinting: Recommendations for a Standardized Methodology","volume":"31","author":"Cave","year":"1993","journal-title":"J. Clin. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1908","DOI":"10.1128\/JCM.00025-15","article-title":"PhyResSE: A Web Tool Delineating Mycobacterium tuberculosis Antibiotic Resistance and Lineage from Whole-Genome Sequencing Data","volume":"53","author":"Feuerriegel","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"573","DOI":"10.2165\/00003088-200140080-00002","article-title":"Clinical pharmacokinetics of sirolimus","volume":"40","author":"Mahalati","year":"2001","journal-title":"Clin. Pharmacokinet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"666","DOI":"10.1097\/01.ftd.0000175911.70172.2e","article-title":"The role of therapeutic monitoring of everolimus in solid organ transplantation","volume":"27","author":"Mabasa","year":"2005","journal-title":"Ther. Drug Monit."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/12\/1\/171\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:05:51Z","timestamp":1760119551000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/12\/1\/171"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,13]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["antibiotics12010171"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics12010171","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,13]]}}}