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Objective— Reverse cholesterol transport comprises cholesterol efflux from ABCA1-expressing macrophages to apolipoprotein (apo) AI, giving nascent high-density lipoprotein (nHDL), esterification of nHDL-free cholesterol (FC), selective hepatic extraction of HDL lipids, and hepatic conversion of HDL cholesterol to bile salts, which are excreted. We tested this model by identifying the fates of nHDL-[3 H]FC, [14 C] phospholipid (PL), and [125 I]apo AI in serum in vitro and in vivo.Approach and Results— During in vitro incubation of human serum, nHDL-[3 H]FC and [14 C]PL rapidly transfer to HDL and low-density lipoproteins (t 1/2 =2–7 minutes), whereas nHDL-[125 I]apo AI transfers solely to HDL (t 1/2 &lt;10 minutes) and to the lipid-free form (t 1/2 &gt;480 minutes). After injection into mice, nHDL-[3 H]FC and [14 C]PL rapidly transfer to liver (t 1/2 =≈2–3 minutes), whereas apo AI clears witht 1/2 =≈460 minutes. The plasma nHDL-[3 H]FC esterification rate is slow (0.46%/h) compared with hepatic uptake. PL transfer protein enhances nHDL-[14 C]PL but not nHDL-[3 H]FC transfer to cultured Huh7 hepatocytes.Conclusions— nHDL-FC, PL, and apo AI enter different pathways in vivo. Most nHDL-[3 H]FC and [14 C]PL are rapidly extracted by the liver via SR-B1 (scavenger receptor class B member 1) and spontaneous transfer; hepatic PL uptake is promoted by PL transfer protein. nHDL-[125 I]apo AI transfers to HDL and to the lipid-free form that can be recycled to nHDL formation. Cholesterol esterification by lecithin:cholesterol acyltransferase is a minor process in nHDL metabolism. These findings could guide the design of therapies that better mobilize peripheral tissue-FC to hepatic disposal.

ABCA1-Derived Nascent High-Density Lipoprotein–Apolipoprotein AI and Lipids Metabolically Segregate