High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery

ABSTRACT Micro- and nanoscale technologies have radically transformed biological research from genomics to tissue engineering, with the relative exception of microbial cell culture, which is still largely performed in microtiter plates and petri dishes. Here, we present nanoscale culture of the oppo...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Anand Srinivasan, Kai P. Leung, Jose L. Lopez-Ribot, Anand K. Ramasubramanian
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2013
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/2d7fa4b6d08340b59833b8f6b6e19360
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2d7fa4b6d08340b59833b8f6b6e19360
record_format	dspace
spelling	oai:doaj.org-article:2d7fa4b6d08340b59833b8f6b6e193602021-11-15T15:43:09ZHigh-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery10.1128/mBio.00331-132150-7511https://doaj.org/article/2d7fa4b6d08340b59833b8f6b6e193602013-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00331-13https://doaj.org/toc/2150-7511ABSTRACT Micro- and nanoscale technologies have radically transformed biological research from genomics to tissue engineering, with the relative exception of microbial cell culture, which is still largely performed in microtiter plates and petri dishes. Here, we present nanoscale culture of the opportunistic fungal pathogen Candida albicans on a microarray platform. The microarray consists of 1,200 individual cultures of 30 nl of C. albicans biofilms (“nano-biofilms”) encapsulated in an inert alginate matrix. We demonstrate that these nano-biofilms are similar to conventional macroscopic biofilms in their morphological, architectural, growth, and phenotypic characteristics. We also demonstrate that the nano-biofilm microarray is a robust and efficient tool for accelerating the drug discovery process: (i) combinatorial screening against a collection of 28 antifungal compounds in the presence of immunosuppressant FK506 (tacrolimus) identified six drugs that showed synergistic antifungal activity, and (ii) screening against the NCI challenge set small-molecule library identified three heretofore-unknown hits. This cell-based microarray platform allows for miniaturization of microbial cell culture and is fully compatible with other high-throughput screening technologies. IMPORTANCE Microorganisms are typically still grown in petri dishes, test tubes, and Erlenmeyer flasks in spite of the latest advances in miniaturization that have benefitted other allied research fields, including genomics and proteomics. Culturing microorganisms in small scale can be particularly valuable in cutting down time, cost, and reagent usage. This paper describes the development, characterization, and application of nanoscale culture of an opportunistic fungal pathogen, Candida albicans. Despite a more than 2,000-fold reduction in volume, the growth characteristics and drug response profiles obtained from the nanoscale cultures were comparable to the industry standards. The platform also enabled rapid identification of new drug candidates that were effective against C. albicans biofilms, which are a major cause of mortality in hospital-acquired infections.Anand SrinivasanKai P. LeungJose L. Lopez-RibotAnand K. RamasubramanianAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 4, Iss 4 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Anand Srinivasan Kai P. Leung Jose L. Lopez-Ribot Anand K. Ramasubramanian High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery
description	ABSTRACT Micro- and nanoscale technologies have radically transformed biological research from genomics to tissue engineering, with the relative exception of microbial cell culture, which is still largely performed in microtiter plates and petri dishes. Here, we present nanoscale culture of the opportunistic fungal pathogen Candida albicans on a microarray platform. The microarray consists of 1,200 individual cultures of 30 nl of C. albicans biofilms (“nano-biofilms”) encapsulated in an inert alginate matrix. We demonstrate that these nano-biofilms are similar to conventional macroscopic biofilms in their morphological, architectural, growth, and phenotypic characteristics. We also demonstrate that the nano-biofilm microarray is a robust and efficient tool for accelerating the drug discovery process: (i) combinatorial screening against a collection of 28 antifungal compounds in the presence of immunosuppressant FK506 (tacrolimus) identified six drugs that showed synergistic antifungal activity, and (ii) screening against the NCI challenge set small-molecule library identified three heretofore-unknown hits. This cell-based microarray platform allows for miniaturization of microbial cell culture and is fully compatible with other high-throughput screening technologies. IMPORTANCE Microorganisms are typically still grown in petri dishes, test tubes, and Erlenmeyer flasks in spite of the latest advances in miniaturization that have benefitted other allied research fields, including genomics and proteomics. Culturing microorganisms in small scale can be particularly valuable in cutting down time, cost, and reagent usage. This paper describes the development, characterization, and application of nanoscale culture of an opportunistic fungal pathogen, Candida albicans. Despite a more than 2,000-fold reduction in volume, the growth characteristics and drug response profiles obtained from the nanoscale cultures were comparable to the industry standards. The platform also enabled rapid identification of new drug candidates that were effective against C. albicans biofilms, which are a major cause of mortality in hospital-acquired infections.
format	article
author	Anand Srinivasan Kai P. Leung Jose L. Lopez-Ribot Anand K. Ramasubramanian
author_facet	Anand Srinivasan Kai P. Leung Jose L. Lopez-Ribot Anand K. Ramasubramanian
author_sort	Anand Srinivasan
title	High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery
title_short	High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery
title_full	High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery
title_fullStr	High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery
title_full_unstemmed	High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery
title_sort	high-throughput nano-biofilm microarray for antifungal drug discovery
publisher	American Society for Microbiology
publishDate	2013
url	https://doaj.org/article/2d7fa4b6d08340b59833b8f6b6e19360
work_keys_str_mv	AT anandsrinivasan highthroughputnanobiofilmmicroarrayforantifungaldrugdiscovery AT kaipleung highthroughputnanobiofilmmicroarrayforantifungaldrugdiscovery AT josellopezribot highthroughputnanobiofilmmicroarrayforantifungaldrugdiscovery AT anandkramasubramanian highthroughputnanobiofilmmicroarrayforantifungaldrugdiscovery
_version_	1718427575215194112

High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery

Ejemplares similares