Of special note Abcg g has been identified

Of special note, Abcg5/g8 has been identified as Lith9 by QTL studies in mice. As shown in Fig. 4, Lith9 is localized on mouse chromosome 17 and is co-localized with a genetic biomarker D17Mit155 at approximately 55 centimorgans (cM). In the Lith9 QTL region, Abcg5/g8 is a strong candidate for this gallstone gene. Subsequently, ABCG5/G8 is found to be associated with gallstones in patients (human LITH9). Furthermore, many research groups reported that two gallstone-associated variants in ABCG5/G8, specifically ABCG5-R50C and ABCG8-D19H, are involved in the pathogenesis of gallstones not only in Germans and Chileans, but also in Chinese and Indians. These studies strongly suggest that ABCG5-R50C and ABCG8-D19H may play a crucial role in hepatic cholesterol hypersecretion, thus leading to the formation of cholesterol-supersaturated bile in humans. Sitosterolemia is caused by a mutation in either the ABCG5 or the ABCG8 gene alone, but not in both simultaneously, and hepatic cholesterol secretion is reduced, but not completely eliminated in these patients. To explore the mechanism underlying the effect of ABCG5/G8 on biliary sterol secretion, biliary cholesterol and sitostanol secretion is quantified for 6 h in Abcg8 knockout mice. Mass transport rate of [3H]sitostanol from plasma HDL into bile is significantly faster than that of [14C]cholesterol in wild-type mice; however, reduced amounts of [14C]cholesterol and no [3H]sitostanol are detected in bile of Abcg8 knockout mice. These results clearly demonstrate that the TPMPA of the Abcg8 gene alone significantly reduces, but does not eliminate hepatic cholesterol secretion. In addition, biliary cholesterol studies found that hepatic cholesterol output is significantly reduced, but cholesterol is still secreted into bile in mice with the deletion of either Abcg5 or Abcg8 alone, or both. Consistent with the human results, these mouse data strongly suggest that an ABCG5/G8-independent pathway could also be involved in regulating hepatic cholesterol secretion in humans and mice. Thus, it needs to be further investigated whether disruption of the Abcg5/g8 genes or the Abcg8 gene alone protects against the formation of cholesterol gallstones in gallstone-susceptible C57BL/6J mice fed a lithogenic diet for 8 weeks. It is surprising to find that although the prevalence of gallstones is significantly reduced in Abag5/g8 double knockout and Abag8 knockout mice, classical parallelogram-shaped cholesterol monohydrate crystals and gallstones are still found in these mice during the 8-week period of the lithogenic diet feeding. In addition, these studies provided clear evidence showing that (i) the ABCG5/G8-independent pathway accounts for 30%–40% of hepatic cholesterol output in the lithogenic state and has an effect on regulating biliary secretion of cholesterol in response to high dietary cholesterol; (ii) in the absence of ABCG5/G8, it plays a pivotal role in biliary cholesterol secretion and the pathogenesis of cholesterol gallstones; (iii) it is able to regulate hepatic secretion of HDL-derived cholesterol, but not sitostanol; and (iv) its activity in the liver is not regulated by the LXR agonist through the LXR signaling pathway. These results support a novel concept that the ABCG5/G8-independent pathway is essential for regulating hepatic cholesterol secretion in the absence of ABCG5/G8 and also plays a determinant role in gallstone formation in mice. Although biliary phospholipids are possibly derived from the cell membranes of hepatocytes, their compositions differ significantly. The cell membranes of hepatocytes contain high levels of phosphatidylcholine (such as lecithin), phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, and sphingomyelin. The major source of phosphatidylcholine molecules destined for secretion into bile is hepatic synthesis. However, a fraction of biliary phosphatidylcholines may also originate from the surface phospholipid coat of HDL particles. A P-glycoprotein member of the multi-drug resistance gene family, ABCB4 plays an important role in regulating hepatic secretion of biliary phospholipids because the deletion of the Abcb4 gene results in a complete inhibition of biliary phospholipid secretion in mice. ABCB4 may be responsible for the translocation or “flip” of phosphatidylcholines from the endoplasmic (inner) to ectoplasmic (outer) leaflet of the canalicular membrane bilayer, and the action of ABCB4 may form phosphatidylcholine-rich microdomains within the outer membrane leaflet. Furthermore, the mutation of the ABCB4 gene in humans is the molecular defect underlying progressive familial intrahepatic cholestasis, type 3 (PFIC3). Biliary phospholipids also play a key role in solubilizing excess cholesterol in vesicles. Low phospholipid-associated cholelithiasis (LPAC) is characterized mainly by the occurrence of intrahepatic and gallbladder microlithiasis in young adults associated with ABCB4 mutations. The Abcb4 knockout mouse is an excellent model for studying the pathogenesis of LPAC. Even on a chow diet, Abcb4 knockout mice spontaneously develop gallstones that are composed mainly of needle-shaped anhydrous cholesterol crystals, which form in phospholipid-deficient gallbladder bile with its relative biliary lipid composition that is in the far-left crystallization region of the phase diagram. These studies support the concept that this gene is a monogenic risk factor for this “peculiar” form of cholesterol gallstones and a target for novel therapeutic strategies.