Targeting Primary Cilia Immune Receptor Proteins for the Treatment of Polycystic Kidney Disease Mechanisms
Background: Primary cilia are cellular organelles project from the cell surface of mammalian cell and play important roles in vertebrate development, organogenesis, health, and others genetic diseases. Primary cilium functions as a mechano-sensor and chemo-sensor. Defect in primary cilia causes the progression of polycystic kidney disease (PKD) which further leads to the inflammatory responses. We, therefore, investigated the role of Toll-like receptors 4 and 9 (TLR) in primary cilia towards PKD. Purpose: The main purpose of the proposed study is to identify and target the immune reactive proteins i.e. TLRs in the primary cilia. By targeting those primary cilia immune reactive proteins using suitable agonist and antagonists to study the control of cystic formation and their progression mechanisms. Methods: To target the ciliary immune TLR proteins (TLR4 and TLR9), we did immunostaining to evaluate their localization on primary cilia. Cilia lengths were measured and compared using differential interference contrast (DIC) and fluorescent imaging techniques. The in vitro3D cyst progression was monitored by adding agonists lipopolysaccharide (LPS) and oligodeoxynucleotides (ODN) and antagonist 4-hydroxy chloroquine (HCQ). Results: From our results we found that the TLR antagonist HCQ increases ciliary length in treated scrambled control, Pkd2knockout (KO) and TLR4KOcells as an immune response, whereas opposite results were observed with TLR9KO. However, the selected agonists for TLRs (LPS/ODN) increases cilia length in TLR9KO cells and decreases scrambled control, Pkd2KO and TLR4KO. In our 3D cyst cultures, we used agonists and antagonist for both the TLRs and observed that the cyst formations and progressions were inversely related to the cilia lengths. From these observations, we speculated that the new ciliary TLR proteins have a role in cystic progression. In conclusion, we found that the TLRs agonists/antagonist can modulate cilia length and TLRs role in inflammatory actions. The primary cilium already has central roles throughout cell biology, but here we propose, for the first time, that the cilium and the regulation of its structural importance in inflammation of PKD.
Other Pharmacy and Pharmaceutical Sciences, Pharmacy and Pharmaceutical Sciences
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