Lysosomal storage disease enzymes
The present invention provides compositions of recombinant human lysosomal acid lipase having particular glycosylation patterns for internalization into target cells, a vector containing the nucleic acid encoding human lysosomal acid lipase, a host cell transformed with the vector, pharmaceutical compositions comprising the recombinant human lysosomal acid lipase and method of treating conditions associated with lysosomal acid lipase deficiency.
1. A nucleic acid vector comprising a nucleic acid sequence encoding human recombinant lysosomal acid lipase comprising the amino acid sequence of SEQ ID NO: 2, 3, 4, or 19, wherein the nucleic acid sequence is operably linked to a promoter and wherein the nucleic acid sequence comprises an adenine (A) at position 93 relative to SEQ ID NO: 5.
2. An isolated host cell transformed with a vector according to claim 1 .
3. The vector of claim 1 , wherein the vector is configured for expression in a cell of:
(a) an avian;
(b) a mammal;
(c) a bacteria; or
(d) a plant.
4. The vector of claim 3 , wherein
the avian is a chicken, turkey, or quail.
5. The vector of claim 3 , wherein the avian cell is an oviduct cell.
6. The vector of claim 5 , wherein the oviduct cell is a tubular gland cell.
7. The vector of claim 1 , wherein the vector is a retroviral vector or a lentiviral vector.
8. The vector of claim 7 , wherein the retroviral vector is a murine leukemia virus (MLV) vector.
9. The vector of claim 1 , wherein the promoter comprises at least a portion of a cytomegalovirus (CMV) promoter, a rous-sarcoma virus (RSV) promoter, a β-actin promoter, a murine leukemia virus (MLV) promoter, a mouse mammary tumor virus (MMTV) promoter, an ovalbumin promoter, a lysozyme promoter, a conalbumin promoter, an ovomucoid promoter, an ovomucin promoter, or an ovotransferrin promoter, or wherein the promoter comprises an ovomucoid and ovotransferrin (MDOT) promoter.
10. The vector of claim 9 , wherein the vector further comprises:
(a) a replication origin sequence;
(b) an enhancer sequence;
(c) a long terminal repeat (LTR) sequence;
(d) a 5′ untranslated region (UTR);
(e) an internal ribosome entry site (IRES) element;
(f) a signal peptide coding sequence operably linked to the human recombinant lysosomal acid lipase;
(g) a 3′ UTR;
(h) a polyadenylation sequence; or
(i) a selectable marker sequence.
11. A composition comprising the vector of claim 1 .
12. A viral particle comprising the vector of claim 1 .
13. The nucleic acid vector of claim 1 , wherein the nucleic acid sequence of the human lysosomal acid lipase has the sequence of nucleotides 5505-6704 of SEQ ID NO: 6.