US-12618059-B2 - Anc80 encoding sphingolipid-metabolizing proteins

Abstract

The present disclosure pertains to the use of an Anc80 viral vector that encodes a sphingolipid-metabolizing protein such as acid ceramidase to achieve expression of the sphingolipid-metabolizing protein in a mammalian cell or group of cells. Expression of the protein from the Anc80 vector reduces high levels of ceramide in the cell that lead to cell death or senescence.

Inventors

• Efrat ELIYAHU
• Adam VINCEK
• Michael Katz

Assignees

• ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI

Dates

Publication Date

20260505

Application Date

20190307

Claims (5)

1. 1 . A method to restore heart function in a subject following ischemia, reperfusion injury or myocardial infarction (MI), the method comprising: directly administering a therapeutically effective amount of an Anc80 viral vector comprising a nucleotide sequence encoding an acid ceramidase protein to cardiac cells of the subject in vivo, wherein the nucleotide sequence encoding the acid ceramidase is operatively linked to an expression control sequence.
2. 2 . The method of claim 1 , wherein the nucleotide sequence encoding the acid ceramidase is as set out in one of SEQ ID NO: 1, SEQ ID NO: 6 and SEQ ID NO: 7.
3. 3 . The method of claim 1 , wherein the nucleotide sequence encoding the acid ceramidase is as set out in SEQ ID NO: 1.
4. 4 . The method of claim 1 , wherein the nucleotide sequence encoding the acid ceramidase is as set out in SEQ ID NO: 6.
5. 5 . The method of claim 1 , wherein the nucleotide sequence encoding the acid ceramidase is as set out in SEQ ID NO: 7.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a national stage application under 35 U.S.C. § 371 of International Application No. PCT/US2019/021201, filed Mar. 7, 2019, and published as WO 2020/005341 on Jan. 2, 2020, which claims the benefit of U.S. provisional application No. 62/692,185, filed Jun. 29, 2018. The entire contents of each of the prior applications are hereby incorporated by reference in their entirety. SEQUENCE LISTING The instant application contains a Sequence Listing, created on Dec. 18, 2018; the file, in ASCII format, is designated 3710047P_SequenceListing_ST25.txt and is 39.9 kilobytes in size. The file is hereby incorporated by reference in its entirety into the instant application. TECHNICAL FIELD The present disclosure relates generally to the use of sphingolipid-metabolizing proteins to improve the survival of mammalian cells. Exposure to sphingolipid metabolizing proteins such as acid ceramidase protein or expression of sphingolipid metabolizing proteins from an Anc80 vector inhibits cell death and/or senescence, preserves and restores normal cellular function, and prolongs survival of cells. BACKGROUND OF THE DISCLOSURE Ceramides are bioactive lipid mediators that influence cell proliferation, differentiation, adhesion and migration. These important cellular lipids are involved in signal transduction pathways such as cell death, senescence and the biosynthesis of other complex sphingolipids There are several studies that support the association of ceramide with cellular and organismal aging. High levels of cellular ceramides can trigger cell death or senescence while ceramide metabolites, such as ceramide-1-phosphate and sphingosine-1-phosphate (S1P), are associated with cell survival and proliferation. In acute myocardial infarction (MI), for example, the level of lipids in the patient's blood can serve to predict the risk for complication. In particular, high levels of ceramides have been associated with a higher probability of recurring events and mortality. Therefore, the ability to moderate ceramide to inhibit cell death and to prevent or reverse senescence may be important therapeutically to promote normal cell function and survival. Though there are several pathways to synthesize ceramide, there is only one way to physiologically hydrolyze it; ceramide can be hydrolyzed into sphingosine by active ceramidase enzymes, including acid ceramidase (AC). The hydrolysis of ceramide produces sphingosine, which is rapidly converted to S1P, a “pro-survival” lipid. We have previously shown that administration of recombinant acid ceramidase (AC) (see U.S. Pat. No. 8,961,962 to Schuchman et al., herein incorporated by reference) promotes ex vivo survival of cells. However, what is needed is a gene delivery method that achieves long-term expression of a sphingolipid-metabolizing enzyme in mammalian cells in vivo to inhibit cell death and senescence and initiate survival. SUMMARY OF THE DISCLOSURE The present disclosure provides a method for promoting survival and restoring function of cells or tissue in vitro or in vivo by administration of a viral vector that encodes a sphingolipid-metabolizing protein, and/or of a protein. In one embodiment, the disclosure relates to a method for promoting survival and restoring function of cells or tissue in vivo by administration of a sphingolipid-metabolizing protein directly to the cell, cells or tissue. In a related aspect, the disclosure relates to a method to preserve and restore function to cardiac cells following ischemia, reperfusion injury, myocardial infarction (MI), pulmonary arterial hypertension (PAH), or other stress-related events comprising contacting said cardiac cells in vivo with a sphingolipid-metabolizing protein or an Anc80 viral vector encoding a sphingolipid-metabolizing protein. In a related aspect, the disclosure relates to a method for preserving and/or restoring heart function in a subject following MI, the method comprising administering to the subject a therapeutically effective amount of a sphingolipid-metabolizing protein, an Anc80 viral vector encoding a sphingolipid-metabolizing protein, or a combination thereof. The sphingolipid-metabolizing protein is selected from the group consisting of (1) a ceramidase; (2) sphingosine kinase (SPHK); (3) sphingosine-1-phosphate receptor (SIPR); (4) ceramidase kinase (CERK) or a combination of (1), (2), (3), and (4). In one embodiment, the sphingolipid-metabolizing protein is a ceramidase. In one embodiment the sphingolipid-metabolizing protein is an acid ceramidase. In one embodiment, the sphingolipid-metabolizing protein is a neutral ceramidase. In yet another embodiment, the sphingolipid-metabolizing protein is an alkaline ceramidase. In one embodiment, ceramidase is encoded by a nucleic acid selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12. In yet another as