Genotypic, proteomic, and phenotypic approaches to decipher the response to caspofungin and calcineurin inhibitors in clinical isolates of echinocandin-resistant Candida glabrata.
Genotypic, proteomic, and phenotypic approaches to decipher the response to caspofungin and calcineurin inhibitors in clinical isolates of echinocandin-resistant Candida glabrata.
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DOI：
10.1093/jac/dkab454
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发表时间：
2022-02-23
期刊：
影响因子：
--
通讯作者：
Parra-Giraldo CM
中科院分区：
文献类型：
--
作者：
Ceballos-Garzon A;Monteoliva L;Gil C;Alvarez-Moreno C;Vega-Vela NE;Engelthaler DM;Bowers J;Le Pape P;Parra-Giraldo CM
棘白菌素耐药性是一个很大的问题，因为这些药物被推荐作为侵袭性念珠菌病的一线治疗。棘白菌素耐药性是由FKS基因突变引起的。然而，途径对于实现耐受性、进化和维持抗性至关重要。因此，了解卡泊芬净反应中涉及的生物学过程和蛋白质可能提供新的治疗靶点的线索。我们确定了耐药机制，并评估了蛋白质组对卡泊芬净暴露的反应。然后，我们评估了FK506和头孢菌素A（CsA）对卡泊芬净耐药光滑念珠菌分离株的钙调磷酸酶抑制作用的表型影响。通过NGS分析了25个与卡泊芬净耐药相关的基因，然后研究了对卡泊芬净暴露的定量蛋白质组学反应。然后，在存在FK 506和CsA的情况下进行卡泊芬净的药敏试验。分析了钙调磷酸酶抑制剂/卡泊芬净组合对热应激（40 ° C）、氧化应激（0.2和0.4mM甲萘醌）和生物膜形成（聚氨酯导管）的影响。最后，开发了使用芽生孢子（1 × 109 cfu/mL）的大蜡螟模型，以评价组合对幼虫存活的影响。 FKS2基因中存在F659-del。蛋白质组学研究表明，一些上调表达的蛋白质参与了细胞壁的生物合成、胁迫应答和致病过程，其中一些蛋白质是钙调素-钙调神经磷酸酶途径的成员。因此，探索了钙调蛋白抑制的影响。钙调素抑制恢复卡泊芬净的敏感性，降低能力，以应对压力条件，并减少生物膜的形成和体内致病性。我们的研究结果证实，钙调蛋白-钙调神经磷酸酶-Crz1可以提供一个相关的目标，在危及生命的侵袭性念珠菌病。
Echinocandin resistance represents a great concern, as these drugs are recommended as first-line therapy for invasive candidiasis. Echinocandin resistance is conferred by mutations in FKS genes. Nevertheless, pathways are crucial for enabling tolerance, evolution, and maintenance of resistance. Therefore, understanding the biological processes and proteins involved in the response to caspofungin may provide clues indicating new therapeutic targets. We determined the resistance mechanism and assessed the proteome response to caspofungin exposure. We then evaluated the phenotypic impact of calcineurin inhibition by FK506 and cephalosporine A (CsA) on caspofungin-resistant Candida glabrata isolates. Twenty-five genes associated with caspofungin resistance were analysed by NGS, followed by studies of the quantitative proteomic response to caspofungin exposure. Then, susceptibility testing of caspofungin in presence of FK506 and CsA was performed. The effects of calcineurin inhibitor/caspofungin combinations on heat stress (40°C), oxidative stress (0.2 and 0.4 mM menadione) and on biofilm formation (polyurethane catheter) were analysed. Finally, a Galleria mellonella model using blastospores (1 × 109 cfu/mL) was developed to evaluate the impact of the combinations on larval survival. F659-del was found in the FKS2 gene of resistant strains. Proteomics data showed some up-regulated proteins are involved in cell-wall biosynthesis, response to stress and pathogenesis, some of them being members of calmodulin–calcineurin pathway. Therefore, the impact of calmodulin inhibition was explored. Calmodulin inhibition restored caspofungin susceptibility, decreased capacity to respond to stress conditions, and reduced biofilm formation and in vivo pathogenicity. Our findings confirm that calmodulin-calcineurin-Crz1 could provide a relevant target in life-threatening invasive candidiasis.
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影响因子：
14.9
作者：
Dobson L;Reményi I;Tusnády GE
通讯作者：
Tusnády GE
影响因子：
4.5
作者：
Alonso-Monge R;Guirao-Abad JP;Sánchez-Fresneda R;Pla J;Yagüe G;Argüelles JC
通讯作者：
Argüelles JC
影响因子：
4.9
作者：
Garcia-Effron, Guillermo;Katiyar, Santosh K.;Perlin, David S.
通讯作者：
Perlin, David S.
DOI：
10.1007/978-1-61779-539-8_33
发表时间：
2012-01-01
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Fallon, John;Kelly, Judy;Kavanagh, Kevin
通讯作者：
Kavanagh, Kevin
影响因子：
--
作者：
de Groot, Piet W. J.;Kraneveld, Eefje A.;Weig, Michael
通讯作者：
Weig, Michael