Sonochemical induction of ABCA1 expression and compositions therefor
First Claim
1. A method for increasing HDL-cholesterol levels in the blood of a subject, the method comprising the steps of:
- (a) intravenously co-administering to the subject a plasmid vector encoding the active form of ATP-binding cassette transporter A1 (ABCA1), and sonochemically-active microspheres;
wherein the vector comprises an expressible open reading frame encoding the active form of ABCA1 and at least one sequence adapted to promote expression of the open reading frame in a mammalian cell; and
wherein the sonochemically-active microspheres comprise gas bubbles encapsulated within shells comprising a protein, a lipid, or a combination thereof, the microspheres being disruptable upon exposure to ultrasonic acoustic energy to release the encapsulated gas bubbles;
(b) ultrasonically imaging a tissue of the subject to be transfected by the plasmid while the plasmid and microspheres are circulating through the vasculature of the tissue and thereby detecting the presence of the microspheres in the vasculature of the tissue; and
(c) while the microspheres are present in the tissue, applying pulses of ultrasonic energy to the tissue at an acoustical energy level higher than a level required for imaging and at a sufficient energy level to disrupt the microspheres and release the gas bubbles therefrom, the pulses of ultrasonic energy and release of gas bubbles thereby temporarily increasing the porosity of cells in the tissue to facilitate entry of the plasmid into the cells;
wherein the plasmid transfects the cells and elicits production of an increased level of HDL-cholesterol in the blood of the subject compared to the HDL-cholesterol level in the blood of the subject prior to administering the plasmid.
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Abstract
The present invention provides methods for transfecting cells (e.g., liver cells) to express ABCA1 and increase HDL in the blood. The compositions described herein comprise a pharmaceutically acceptable aqueous carrier containing sonochemically-active microspheres together with a plasmid DNA construct encoding an active form of ABCA1 and at least one promoter for the expression thereof. Preferably, the sonochemically-active microspheres comprise, consist essentially of, or consist of gas bubbles (e.g., a fluorocarbon gas, such as octafluoropropane) encapsulated within protein-containing or lipid-containing shells (e.g., human serum albumin shells). The microspheres are disruptable by exposure to ultrasonic acoustic energy to release the encapsulated gas.
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Citations
24 Claims
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1. A method for increasing HDL-cholesterol levels in the blood of a subject, the method comprising the steps of:
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(a) intravenously co-administering to the subject a plasmid vector encoding the active form of ATP-binding cassette transporter A1 (ABCA1), and sonochemically-active microspheres;
wherein the vector comprises an expressible open reading frame encoding the active form of ABCA1 and at least one sequence adapted to promote expression of the open reading frame in a mammalian cell; and
wherein the sonochemically-active microspheres comprise gas bubbles encapsulated within shells comprising a protein, a lipid, or a combination thereof, the microspheres being disruptable upon exposure to ultrasonic acoustic energy to release the encapsulated gas bubbles;(b) ultrasonically imaging a tissue of the subject to be transfected by the plasmid while the plasmid and microspheres are circulating through the vasculature of the tissue and thereby detecting the presence of the microspheres in the vasculature of the tissue; and (c) while the microspheres are present in the tissue, applying pulses of ultrasonic energy to the tissue at an acoustical energy level higher than a level required for imaging and at a sufficient energy level to disrupt the microspheres and release the gas bubbles therefrom, the pulses of ultrasonic energy and release of gas bubbles thereby temporarily increasing the porosity of cells in the tissue to facilitate entry of the plasmid into the cells; wherein the plasmid transfects the cells and elicits production of an increased level of HDL-cholesterol in the blood of the subject compared to the HDL-cholesterol level in the blood of the subject prior to administering the plasmid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. A method for increasing HDL-cholesterol levels in the blood of a subject, the method comprising the steps of:
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(a) intravenously administering to the subject a composition comprising a mixture of a nucleic acid encoding an expressible open reading frame and sonochemically-active microspheres in a pharmaceutically acceptable aqueous carrier;
wherein the nucleic acid of the mixture consists of a plasmid vector, the expressible open reading frame consists of an expressible open reading frame encoding the active form of ATP-binding cassette transporter A1 (ABCA1), and the plasmid vector also includes at least one sequence adapted to promote expression of the open reading frame in a mammalian cell; and
wherein the sonochemically-active microspheres comprise gas bubbles encapsulated within shells comprising a protein, a lipid, or a combination thereof, the microspheres being disruptable upon exposure to ultrasonic acoustic energy to release the encapsulated gas bubbles;(b) ultrasonically imaging a tissue of the subject to be transfected by the plasmid vector while the composition is circulating through the vasculature of the tissue and thereby detecting the presence of the microspheres in the vasculature of the tissue; and (c) while the microspheres are present in the tissue, applying pulses of ultrasonic energy to the tissue at an acoustical energy level higher than a level required for imaging and at a sufficient energy level to disrupt the microspheres and release the gas bubbles therefrom, the pulses of ultrasonic energy and release of gas bubbles thereby temporarily increasing the porosity of cells in the tissue to facilitate entry of the plasmid into the cells; wherein the tissue comprises liver tissue, intestinal parenchymal tissue, or a combination thereof;
the plasmid transfects the cells and elicits production of an increased level of HDL-cholesterol in the blood of the subject compared to the HDL-cholesterol level in the blood of the subject prior to administering the composition; and
the pulses are applied at a rate of about 6 to 8 pulses per minute, with pulse durations in the range of about 500 to about 2000 milliseconds per pulse, and at an acoustic energy mechanical index (MI) of about 1 to about 2 MI. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24)
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Specification