Gene therapy for treating familial hypercholesterolemia

1. A method for reducing the need for apheresis in a patient having familial hypercholesterolemia (FH), the method comprising administering to the patient a pharmaceutical composition suitable for peripheral vein infusion in the patient, said pharmaceutical composition comprising a suspension of replication deficient recombinant adeno-associated virus (rAAV) in a formulation buffer, wherein:

(a) the rAAV is AAV8.TBG.hLDLR which comprises an AAV8 capsid and a vector genome comprising an AAV 5′ inverted terminal repeat (ITR), expression cassette, and an AAV 3′ ITR, wherein the expression cassette comprises a nucleic acid sequence encoding a human LDL receptor (hLDLR) operably linked to regulatory sequences, wherein the hLDLR coding sequence comprises nucleic acid sequence of SEQ ID NO: 4, wherein regulatory sequences comprise an intron, two copies of an alpha 1 microglobulin/bikunin enhancer element, a liver specific promoter which is a thyroxine binding globulin (TBG) promoter, and a rabbit beta-globin polyadenylation signal, and wherein said vector genome is packaged in the AAV8 capsid;

(b) the formulation buffer comprises an aqueous solution of phosphate buffered saline and a poloxamer; and

(c) the suspension comprises AAV8.TBG.hLDLR in a Genome Copies (GC) titer of at least 1×10 13 GC/mL of the solution.

2. The method according to claim 1 , wherein the patient has a neutralizing antibody titer of less than or equal to 1:10 against the AAV8 capsid.

3. The method according to claim 2 , wherein the method further comprising co-treating the patient with an immunosuppressive regimen.

4. The method according to claim 3 , wherein the patient has a neutralizing antibody titer of at least 1:5 prior to co-treatment with the immunosuppressive regimen.

5. The method according to claim 1 , wherein the patient has a neutralizing antibody titer of less than or equal to 1:5 against the AAV8 capsid.

6. The method according to claim 1 , wherein the rAAV is AAV8.TBG.hLDLR comprising the vector genome which comprises the nucleic acid sequence of nucleotides 1 to 3947 of SEQ ID NO: 6.

7. The method according to claim 1 , wherein the formulation buffer is 180 mM NaCl, 10 mM Na phosphate, 0.001% poloxamer 188, pH 7.3.

8. The method according to claim 1 , wherein the pharmaceutical composition is administered to the patient via a peripheral vein by infusion of a suspension of replication deficient recombinant adeno-associated virus (rAAV) at a dose of (i) at least 5×10 11 Genome Copies (GC)/kg or (ii) 2.5×10 12 GC/kg to 7.5×10 12 GC/kg body weight of the patient as determined by optimized qPCR (oqPCR) or digital droplet PCR (ddPCR).

9. The method according to claim 1 , wherein the patient has been diagnosed with Homozygous FH (HoFH).

10. The method according to claim 1 , wherein the patient has been diagnosed with Heterozygous FH (HeFH).