Multifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells

ABSTRACT Fungal infections cause significant mortality and morbidity worldwide, and the limited existing antifungal reservoir is further weakened by the emergence of strains resistant to echinocandins, a first line of antifungal therapy. Candida glabrata is an opportunistic fungal pathogen that rapi...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Rocio Garcia-Rubio, Cristina Jimenez-Ortigosa, Lucius DeGregorio, Christopher Quinteros, Erika Shor, David S. Perlin
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2021
Materias:	Candida glabrata echinocandins transcriptomics mitochondria antifungal drug tolerance Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/0355009511bd4f31a5ba13af1c8e147f
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:0355009511bd4f31a5ba13af1c8e147f
record_format	dspace
spelling	oai:doaj.org-article:0355009511bd4f31a5ba13af1c8e147f2021-11-10T18:37:52ZMultifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells10.1128/mBio.01959-212150-7511https://doaj.org/article/0355009511bd4f31a5ba13af1c8e147f2021-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01959-21https://doaj.org/toc/2150-7511ABSTRACT Fungal infections cause significant mortality and morbidity worldwide, and the limited existing antifungal reservoir is further weakened by the emergence of strains resistant to echinocandins, a first line of antifungal therapy. Candida glabrata is an opportunistic fungal pathogen that rapidly develops mutations in the echinocandin drug target β-1,3-glucan synthase (GS), which are associated with drug resistance and clinical failure. Although echinocandins are considered fungicidal in Candida sp., a subset of C. glabrata cells survive echinocandin exposure, forming a drug-tolerant cell reservoir, from which resistant mutations are thought to emerge. Despite their importance, the physiology of rare drug-tolerant cells is poorly understood. We used fluorescence-activated cell sorting to enrich for echinocandin-tolerant cells, followed by modified single-cell RNA sequencing to examine their transcriptional landscape. This analysis identified a transcriptional signature distinct from the stereotypical yeast environmental stress response and characterized by upregulation of pathways involved in chromosome structure and DNA topology and downregulation of oxidative stress responses, of which the latter was observed despite increased levels of reactive oxygen species. Further analyses implicated mitochondria in echinocandin tolerance, wherein inhibitors of mitochondrial complexes I and IV reduced echinocandin-mediated cell killing, but mutants lacking various mitochondrial components all showed an echinocandin hypotolerant phenotype. Finally, GS enzyme complexes purified from mitochondrial mutants exhibited normal in vitro inhibition kinetics, indicating that mitochondrial defects influence cell survival downstream of the drug-target interaction. Together, these results provide new insights into the C. glabrata response to echinocandins and reveal a multifactorial role of mitochondria in echinocandin tolerance. IMPORTANCE Echinocandin drugs are a first-line therapy to treat invasive candidiasis, which is a major source of morbidity and mortality worldwide. The opportunistic fungal pathogen Candida glabrata is a prominent bloodstream fungal pathogen, and it is notable for rapidly developing echinocandin-resistant strains associated with clinical failure. Echinocandin resistance is thought to emerge within a small echinocandin-tolerant subset of C. glabrata cells that are not killed by drug exposure, but mechanisms underlying echinocandin tolerance are still unknown. Here, we describe the unique transcriptional signature of echinocandin-tolerant cells and the results of follow-up analyses, which reveal a multifactorial role of mitochondria in C. glabrata echinocandin tolerance. In particular, although chemical inhibition of respiratory chain enzymes increased echinocandin tolerance, deletion of multiple mitochondrial components made C. glabrata cells hypotolerant to echinocandins. Together, these results provide new insights into the C. glabrata response to echinocandins and reveal the involvement of mitochondria in echinocandin tolerance.Rocio Garcia-RubioCristina Jimenez-OrtigosaLucius DeGregorioChristopher QuinterosErika ShorDavid S. PerlinAmerican Society for MicrobiologyarticleCandida glabrataechinocandinstranscriptomicsmitochondriaantifungal drug toleranceMicrobiologyQR1-502ENmBio, Vol 12, Iss 4 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Candida glabrata echinocandins transcriptomics mitochondria antifungal drug tolerance Microbiology QR1-502
spellingShingle	Candida glabrata echinocandins transcriptomics mitochondria antifungal drug tolerance Microbiology QR1-502 Rocio Garcia-Rubio Cristina Jimenez-Ortigosa Lucius DeGregorio Christopher Quinteros Erika Shor David S. Perlin Multifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells
description	ABSTRACT Fungal infections cause significant mortality and morbidity worldwide, and the limited existing antifungal reservoir is further weakened by the emergence of strains resistant to echinocandins, a first line of antifungal therapy. Candida glabrata is an opportunistic fungal pathogen that rapidly develops mutations in the echinocandin drug target β-1,3-glucan synthase (GS), which are associated with drug resistance and clinical failure. Although echinocandins are considered fungicidal in Candida sp., a subset of C. glabrata cells survive echinocandin exposure, forming a drug-tolerant cell reservoir, from which resistant mutations are thought to emerge. Despite their importance, the physiology of rare drug-tolerant cells is poorly understood. We used fluorescence-activated cell sorting to enrich for echinocandin-tolerant cells, followed by modified single-cell RNA sequencing to examine their transcriptional landscape. This analysis identified a transcriptional signature distinct from the stereotypical yeast environmental stress response and characterized by upregulation of pathways involved in chromosome structure and DNA topology and downregulation of oxidative stress responses, of which the latter was observed despite increased levels of reactive oxygen species. Further analyses implicated mitochondria in echinocandin tolerance, wherein inhibitors of mitochondrial complexes I and IV reduced echinocandin-mediated cell killing, but mutants lacking various mitochondrial components all showed an echinocandin hypotolerant phenotype. Finally, GS enzyme complexes purified from mitochondrial mutants exhibited normal in vitro inhibition kinetics, indicating that mitochondrial defects influence cell survival downstream of the drug-target interaction. Together, these results provide new insights into the C. glabrata response to echinocandins and reveal a multifactorial role of mitochondria in echinocandin tolerance. IMPORTANCE Echinocandin drugs are a first-line therapy to treat invasive candidiasis, which is a major source of morbidity and mortality worldwide. The opportunistic fungal pathogen Candida glabrata is a prominent bloodstream fungal pathogen, and it is notable for rapidly developing echinocandin-resistant strains associated with clinical failure. Echinocandin resistance is thought to emerge within a small echinocandin-tolerant subset of C. glabrata cells that are not killed by drug exposure, but mechanisms underlying echinocandin tolerance are still unknown. Here, we describe the unique transcriptional signature of echinocandin-tolerant cells and the results of follow-up analyses, which reveal a multifactorial role of mitochondria in C. glabrata echinocandin tolerance. In particular, although chemical inhibition of respiratory chain enzymes increased echinocandin tolerance, deletion of multiple mitochondrial components made C. glabrata cells hypotolerant to echinocandins. Together, these results provide new insights into the C. glabrata response to echinocandins and reveal the involvement of mitochondria in echinocandin tolerance.
format	article
author	Rocio Garcia-Rubio Cristina Jimenez-Ortigosa Lucius DeGregorio Christopher Quinteros Erika Shor David S. Perlin
author_facet	Rocio Garcia-Rubio Cristina Jimenez-Ortigosa Lucius DeGregorio Christopher Quinteros Erika Shor David S. Perlin
author_sort	Rocio Garcia-Rubio
title	Multifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells
title_short	Multifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells
title_full	Multifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells
title_fullStr	Multifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells
title_full_unstemmed	Multifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells
title_sort	multifactorial role of mitochondria in echinocandin tolerance revealed by transcriptome analysis of drug-tolerant cells
publisher	American Society for Microbiology
publishDate	2021
url	https://doaj.org/article/0355009511bd4f31a5ba13af1c8e147f
work_keys_str_mv	AT rociogarciarubio multifactorialroleofmitochondriainechinocandintolerancerevealedbytranscriptomeanalysisofdrugtolerantcells AT cristinajimenezortigosa multifactorialroleofmitochondriainechinocandintolerancerevealedbytranscriptomeanalysisofdrugtolerantcells AT luciusdegregorio multifactorialroleofmitochondriainechinocandintolerancerevealedbytranscriptomeanalysisofdrugtolerantcells AT christopherquinteros multifactorialroleofmitochondriainechinocandintolerancerevealedbytranscriptomeanalysisofdrugtolerantcells AT erikashor multifactorialroleofmitochondriainechinocandintolerancerevealedbytranscriptomeanalysisofdrugtolerantcells AT davidsperlin multifactorialroleofmitochondriainechinocandintolerancerevealedbytranscriptomeanalysisofdrugtolerantcells
_version_	1718439742292361216

Multifactorial Role of Mitochondria in Echinocandin Tolerance Revealed by Transcriptome Analysis of Drug-Tolerant Cells

Ejemplares similares