Gupta Hrishikesh, Gupta Payal, Kairamkonda Manikyaprabhu, Poluri Krishna Mohan
Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee (IIT-Roorkee) Roorkee-247667 Uttarakhand India
Centre for Nanotechnology, Indian Institute of Technology Roorkee Roorkee-247667 Uttarakhand India.
RSC Adv. 2022 Jun 14;12(27):17570-17584. doi: 10.1039/d2ra02092k. eCollection 2022 Jun 7.
Prevalence of drug resistant strains in hospitalized immune-compromised patients with invasive fungal infections has increased at an unexpected pace. This has greatly pushed researchers in identification of mutations/variations in clinical isolates for better assessment of the prevailing drug resistance trends and also for updating of antifungal therapy regime. In the present investigation, the clinical isolates of were comprehensively characterized at a molecular level using metabolic profiling and transcriptional expression analysis approaches in combination with biochemical, morphological and chemical profiling methods. Biochemically, significant variations in azole susceptibility, surface hydrophobicity, and oxidative stress generation were observed among the isolates as compared to wild-type. The H NMR profiling identified 18 differential metabolites in clinical strains compared to wild-type and were classified into five categories, that include: sugars (7), amino acids and their derivatives (7), nitrogen bases (3) and coenzymes (1). Transcriptional analysis of selective metabolic and regulatory enzymes established that the major differences were found in cell membrane stress, carbohydrate metabolism, amino acid biosynthesis, ergosterol pathway and turnover of nitrogen bases. This detailed molecular level/metabolic fingerprint study is a useful approach for differentiating pathogenic/clinical isolates to that of wild-type. This study comprehensively delineated the differential cellular pathways at a molecular level that have been re-wired by the pathogenic clinical isolates for enhanced pathogenicity and virulence traits.
侵袭性真菌感染的住院免疫功能低下患者中耐药菌株的流行率以意想不到的速度增加。这极大地推动了研究人员对临床分离株中的突变/变异进行鉴定，以便更好地评估当前的耐药趋势，并更新抗真菌治疗方案。在本研究中，结合生化、形态学和化学分析方法，使用代谢谱分析和转录表达分析方法在分子水平上对临床分离株进行了全面表征。在生化方面，与野生型相比，分离株在唑类敏感性、表面疏水性和氧化应激产生方面观察到显著差异。与野生型相比，1H NMR谱分析在临床菌株中鉴定出18种差异代谢物，并分为五类，包括：糖类（7种）、氨基酸及其衍生物（7种）、氮碱基（3种）和辅酶（1种）。对选择性代谢和调节酶的转录分析表明，主要差异存在于细胞膜应激、碳水化合物代谢、氨基酸生物合成、麦角固醇途径和氮碱基周转中。这项详细的分子水平/代谢指纹研究是区分致病性/临床分离株与野生型的有用方法。本研究在分子水平上全面描绘了致病性临床分离株为增强致病性和毒力特征而重新连接的差异细胞途径。
