Laboratório de Estudos Avançados de Microrganismos Emergentes e Resistentes (LEAMER), Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes (IMPG), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
Laboratório de Estudos Avançados de Microrganismos Emergentes e Resistentes (LEAMER), Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes (IMPG), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil; Programa de Pós-Graduação em Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Fungal Biol. 2021 May;125(5):378-388. doi: 10.1016/j.funbio.2020.12.004. Epub 2020 Dec 23.
Candida haemulonii species complex (C. haemulonii, C. haemulonii var. vulnera and Candida duobushaemulonii) is composed by emerging and multidrug-resistant (MDR) yeasts. Candidiasis, the disease caused by these species, is difficult to treat and culminates in clinical failures and patient death. It is well-known that Candida peptidases play important roles in the fungus-host interactions, and hence these enzymes are promising targets for developing new antifungal drugs. Recently, serine-type peptidases were described in clinical isolates of C. haemulonii complex with the ability to cleave relevant key host proteins. Herein, the effects of serine peptidase inhibitors (SPIs) on the cell biology of this fungal complex were evaluated. Initially, eight distinct SPIs (phenylmethylsulfonyl fluoride - PMSF, 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride - AEBSF, N-α-tosyl-l-lysine chloromethyl ketone hydrochloride - TLCK, N-p-tosyl-l-phenylalanine chloromethyl ketone - TPCK, simeprevir, boceprevir, danoprevir and telaprevir) were tested on the fungal growth. TPCK showed the best efficacy in controlling cell proliferation, being selected for the following experiments. This SPI induced changes in the architecture of yeast cells, as observed by scanning electron microscopy, besides injuries at the plasma membrane and reduction in the ergosterol content. TPCK also diminished the ability of yeasts to adhere to abiotic (polystyrene and glass) and biotic (murine macrophages) surfaces in a typically concentration-dependent manner. In addition, the 24 h-treatment of the mature biofilm promoted a decrease in biomass, viability and extracellular matrix. Altogether, our results highlight that SPIs may be promising new therapeutic agents in the treatment of candidiasis caused by emergent, opportunistic and MDR species forming the C. haemulonii complex.
海氏假丝酵母种复合体（C. haemulonii、C. haemulonii var. vulnera 和 Candida duobushaemulonii）由新兴和多药耐药（MDR）酵母组成。由这些物种引起的念珠菌病难以治疗，最终导致临床失败和患者死亡。众所周知，假丝酵母肽酶在真菌-宿主相互作用中发挥着重要作用，因此这些酶是开发新型抗真菌药物的有前途的靶点。最近，在海氏假丝酵母复合体的临床分离株中描述了丝氨酸型肽酶，这些酶具有切割相关关键宿主蛋白的能力。在此，评估了丝氨酸肽酶抑制剂（SPIs）对该真菌复合体细胞生物学的影响。最初，在真菌生长方面测试了 8 种不同的 SPI（苯甲基磺酰氟-PMSF、4-（2-氨基乙基）苯磺酰氟盐酸盐-AEBSF、N-α-甲苯磺酰-L-赖氨酸氯甲基酮盐酸盐-TLCK、N-对甲苯磺酰-L-苯丙氨酸氯甲基酮-TPCK、simeprevir、boceprevir、danoprevir 和 telaprevir）。TPCK 对控制细胞增殖最有效，被选为后续实验的选择。该 SPI 引起酵母细胞结构的变化，如扫描电子显微镜观察到的，除了质膜损伤和麦角固醇含量降低。TPCK 还以典型的浓度依赖性方式降低了酵母在非生物（聚苯乙烯和玻璃）和生物（鼠巨噬细胞）表面的粘附能力。此外，成熟生物膜的 24 小时处理促进了生物量、活力和细胞外基质的减少。总之，我们的结果强调，SPIs 可能是治疗由新兴、机会性和 MDR 物种组成的海氏假丝酵母复合体引起的念珠菌病的有前途的新型治疗剂。
