ATP-binding cassette subfamily G member 2 (ABCG2) is a physiologically important urate transporter. Accumulating evidence demonstrates that congenital dysfunction of ABCG2 is an important genetic risk factor in gout and hyperuricemia; recent studies suggest the clinical significance of both common and rare variants of ABCG2. However, the effects of rare variants of ABCG2 on the risk of such diseases are not fully understood. Here, using a cohort of 250 Czech individuals of European descent (68 primary hyperuricemia patients and 182 primary gout patients), we examined exonic non-synonymous variants of ABCG2. Based on the results of direct sequencing and database information, we experimentally characterized nine rare variants of ABCG2: R147W (rs372192400), T153M (rs753759474), F373C (rs752626614), T421A (rs199854112), T434M (rs769734146), S476P (not annotated), S572R (rs200894058), D620N (rs34783571), and a three-base deletion K360del (rs750972998). Functional analyses of these rare variants revealed a deficiency in the plasma membrane localization of R147W and S572R, lower levels of cellular proteins of T153M and F373C, and null urate uptake function of T434M and S476P. Accordingly, we newly identified six rare variants of ABCG2 that showed lower or null function. Our findings contribute to deepening the understanding of ABCG2-related gout/hyperuricemia risk and the biochemical characteristics of the ABCG2 protein.

Department of Cell Biology Faculty of Science Charles University 128 00 Prague 2 Czech Republic

Department of Integrative Physiology and Bio Nano Medicine National Defense Medical College Saitama 359 8513 Japan

Department of Pediatrics and Adolescent Medicine 1st Faculty of Medicine Charles University and General University Hospital 121 08 Prague 2 Czech Republic

Department of Pharmacy The University of Tokyo Hospital Tokyo 113 8655 Japan

Department of Probability and Mathematical Statistics Faculty of Mathematics and Physics Charles University 121 16 Prague 2 Czech Republic

Institute of Rheumatology 128 50 Prague 2 Czech Republic

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Familial early-onset hyperuricemia and gout associated with a newly identified dysfunctional variant in urate transporter ABCG2