Inhibition of autophagy with Chloroquine enhanced apoptosis induced by 5-aminolevulinic acid-photodynamic therapy in secondary hyperparathyroidism primary cells and organoids

Abstract

Secondary hyperparathyroidism (SHPT), the most common complication in the later stage of chronic kidney disease (CKD), seriously affects quality of life and the survival time of patients. At present, the conventional drugs and surgical methods still cannot fully meet the needs of clinical treatment. It is quite significant to develop effective and minimally invasive treatment methods. 5-Aminolevulinic acid-mediated photodynamic therapy (5-ALA-PDT), an alternative treatment relying on light irradiation, photosensitizer, and oxygen to produce a series of cytotoxic effects on tissue, is a promising technique for treating SHPT. We have successfully cultivated SHPT primary cells and organoids, and further proved that the amount of 5-ALA transformed into protoporphyrin IX in a time- and concentration-dependent manner. Also, 5-ALA-PDT exerted a cytotoxic effect on both primary cells and organoids by the cell counting kit (CCK-8) assay. Mechanically, 5-ALA-PDT increased the number of autophagosomes, and autophagy- and apoptosis-related proteins were upregulated markedly by western-blotting. The autophagy inhibitor Chloroquine (CQ) significantly increased the proportion of apoptotic cells, while the autophagy inducer rapamycin decreased the inhibitory ability of 5-ALA-PDT in SHPT primary cells. In brief, 5-ALA-PDT exhibits a phototoxic effect on SHPT primary cells and organoids. Autophagy and apoptosis are involved in the mechanism, and autophagy plays a role in promoting survival and inhibiting apoptosis. Therefore, the use of autophagy inhibitors can increase the sensitivity of SHPT cells and organoids treated with 5-ALA-PDT.

Keywords: 5-Aminolevulinic acid; Apoptosis; Autophagy; Organoids; Potodynamic therapy; Secondary hyperparathyroidism.