Unidad de Investigación, Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunidad Valenciana (FISABIO), Hospital General Universitario de Elche, Camí de l'Almazara 11, Elche, 03203 Alicante, Spain.
Departamento de Bioquímica y Biología Molecular, Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche (IDiBE), Universidad Miguel Hernández, Avda, Universidad s/n, Ed. Torregaitán, Elche, 03202 Alicante, Spain.
Int J Mol Sci. 2021 Feb 2;22(3):1477. doi: 10.3390/ijms22031477.
D-amino acid oxidase (DAAO) is an enzyme that catalyzes the oxidation of D-amino acids generating HO. The enzymatic chimera formed by DAAO bound to the choline-binding domain of N-acetylmuramoyl-L-alanine amidase (CLytA) induces cytotoxicity in several pancreatic and colorectal carcinoma and glioblastoma cell models. In the current work, we determined whether the effect of CLytA-DAAO immobilized in magnetic nanoparticles, gold nanoparticles, and alginate capsules offered some advantages as compared to the free CLytA-DAAO. Results indicate that the immobilization of CLytA-DAAO in magnetic nanoparticles increases the stability of the enzyme, extending its time of action. Besides, we compared the effect induced by CLytA-DAAO with the direct addition of hydrogen peroxide, demonstrating that the progressive generation of reactive oxygen species by CLytA-DAAO is more effective in inducing cytotoxicity than the direct addition of HO. Furthermore, a pilot study has been initiated in biopsies obtained from pancreatic and colorectal carcinoma and glioblastoma patients to evaluate the expression of the main genes involved in resistance to CLytA-DAAO cytotoxicity. Based on our findings, we propose that CLytA-DAAO immobilized in magnetic nanoparticles could be effective in a high percentage of patients and, therefore, be used as an anti-cancer therapy for pancreatic and colorectal carcinoma and glioblastoma.
D-氨基酸氧化酶（DAAO）是一种能够催化 D-氨基酸氧化生成 HO 的酶。DAAO 与 N-乙酰胞壁酰-L-丙氨酸酰胺酶（CLytA）的胆碱结合域结合形成的酶杂合体在几种胰腺和结直肠癌细胞系和神经胶质瘤细胞模型中诱导细胞毒性。在目前的工作中，我们确定了固定在磁性纳米粒子、金纳米粒子和藻酸盐胶囊中的 CLytA-DAAO 是否比游离的 CLytA-DAAO 具有一些优势。结果表明，CLytA-DAAO 固定在磁性纳米粒子中增加了酶的稳定性，延长了其作用时间。此外，我们比较了 CLytA-DAAO 诱导的效果与直接添加过氧化氢的效果，证明 CLytA-DAAO 逐步生成活性氧比直接添加 HO 更能有效诱导细胞毒性。此外，已经启动了一项针对胰腺和结直肠癌细胞系和神经胶质瘤患者活检的初步研究，以评估参与 CLytA-DAAO 细胞毒性耐药的主要基因的表达。基于我们的发现，我们提出固定在磁性纳米粒子中的 CLytA-DAAO 可能对很大一部分患者有效，因此可用于胰腺和结直肠癌细胞系和神经胶质瘤的抗癌治疗。
