Abstract 2610: Histone deacetylase inhibitors mediate pharmacological rescue of the ABCG2 Q141K variant: Potential for therapeutics in cancer and gout
Authors: Basseville Agnes , Tamaki Akina , Ierano Caterina , Ward Yvona , Robey Robert W. , Hegde Ramanujan , Bates Susan E. 
Published: June 2012

ABCG2 is an ATP-binding cassette half-transporter that has garnered interest in pharmacogenomics as a modulator of oral drug absorption, drug excretion, and drug distribution through expression at the blood-brain barrier and the maternal-fetal barrier. A single nucleotide polymorphism (SNP) C421A in ABCG2, resulting in a glutamic acid to lysine mutation at amino acid 141 (Q141K), confers impaired protein expression and function. Pharmacogenomic studies have linked the SNP to increased exposure to substrate drugs including irinotecan, topotecan, sunitinib and gefitinib. Moreover, it has been shown that 10 % of gout cases can be attributed to the Q141K variant.

We have studied the biology underlying altered expression and function of the ABCG2 variant. Q141K protein was resistant to degradation by Endo H, suggesting that it was fully processed and folded. Q141K ABCG2 expression was increased by the lysosome inhibitor bafilomycin, indicating that some of the protein reached the cell surface, although at reduced levels. But the largest fraction of the Q141K variant was retained in an intracellular compartment where a part was eliminated through the proteasomal pathway while the undegraded ABCG2 accumulated in the aggresomes, as shown by colocalization of the transporter with the centrosome marker, γ-tubulin. The ability of several agents to impact ABCG2 trafficking to the cell surface was evaluated. Mitoxantrone, an ABCG2 substrate previously noted to act as a pharmacological chaperone, induced a 2-fold increase in Q141K ABCG2 expression at the cell surface but only slightly enhanced ABCG2-related efflux. Romidepsin and other histone deacetylase inhibitors (HDIs), known to increase levels of ABCG2 mRNA, were also assayed. A significant increase in total protein, surface expression, and function was seen in Q141K ABCG2 variant when exposed to various HDIs. Immunofluorescence analysis by confocal microscopy showed a dramatic shift to the plasma membrane following exposure to the HDIs, whereas the localization of α-tubulin, HDAC6 and vimentin, involved in aggresome formation and structure, were unchanged. These results support the notion that Q141K ABCG2 variant is trapped in the aggresome and that HDIs aid localization to the surface. Some supplementary experiments showed that the Q141K ABCG2 states of phosphorylation, acetylation and dimerization were unmodified after HDIs treatment, and that the chaperones BiP, Hsp70 or Hsp90 were not involved in rescue. Investigations into the HDI-mediated mechanisms are underway.

In addition to non-oncologic indications, the restoration of ABCG2 function has potential applications in oncology, in improving normal tissue protection and drug elimination, as well as in cancer prevention since several carcinogens are substrates for ABCG2.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2610. doi:10.1158/1538-7445.AM2011-2610