Perfusion circuit and use therein in targeted delivery of macromolecules
A perfusion circuit and a use thereof for delivering a substance to a subject's heart in situ during cardiopulmonary bypass surgery are provided. The perfusion circuit defines a path for re-circulating a solution containing a macromolecular complex through a coronary circulation circuit through a subject's heart during a surgical procedure in which the substance is prevented from being delivered to the subject's other organs.
1. A method of delivering a macromolecular complex to a subject's heart, said method comprising the steps of:
(a) forming a cardiac circuit that isolates a subject's cardiac circulation from the subject's systemic circulation; and
(b) perfusing a first macromolecular complex solution into the cardiac circuit in a retrograde manner, wherein said first macromolecular complex solution comprises a viral vector carrying a target molecule, wherein said viral vector is a recombinant adenoviral (rAd) vector or a recombinant adeno-associated viral (rAAV) vector.
2. The method according to claim 1 , wherein said recombinant adeno-associated viral (rAAV) vector comprises an AAV1 capsid.
3. The method according to claim 1 , wherein said recombinant adeno-associated viral (rAAV) vector comprises an AAV2 capsid.
4. The method according to claim 1 , wherein said recombinant adeno-associated viral (rAAV) vector comprises AAV2 inverted terminal repeat sequences.
5. The method according to claim 1 , wherein said recombinant adeno-associated viral (rAAV) vector comprises a serotype 1 capsid and serotype 2 inverted terminal repeat sequences.
6. The method according to claim 1 , wherein said first macromolecular complex solution is recirculated in the cardiac circuit for up to 30 minutes.
7. The method according to claim 6 , wherein said subject's cardiac circulation is restarted and a second macromolecular complex solution is infused in the circuit for up to 20 minutes.
8. The method according to claim 7 , wherein the second macromolecular complex is different from the first macromolecular complex solution.
9. The method according to claim 1 , wherein the subject's cardiac circulation is stopped for up to 10 minutes, thereby allowing the macromolecular complex solution to dwell.
10. The method according to claim 9 , wherein the subject's cardiac circulation is stopped for up about 5 minutes.
11. The method according to claim 1 , wherein retrograde infusion is through the coronary sinus.
12. The method according to claim 1 , where said first macromolecular complex is perfused at about 50 mm Hg to 100 mm Hg.
13. The method according to claim 12 , where said first macromolecular complex is perfused at about 100 mm Hg.
14. The method according to claim 12 , where said first macromolecular complex is perfused at about 60 to 80 mm Hg.
15. The method according to claim 1 , wherein said target molecule is a transgene, a chemical moiety, or a DNA molecule which directs the transcription of an mRNA molecule or a RNA silencing molecule.
16. The method according to claim 15 , wherein said target molecule is a transgene.
17. The method according to claim 16 , wherein said transgene encodes a sarcoglycan protein, dystrophin, utrophin, a minidystrophin protein, a microdystrophin protein, calpain, Fukutin, Fukutin-related protein, telethonin, laminin, beta adrenergic receptor kinase 1 (bARK1), beta andrenoreceptor kinase c-terminase (βARKct), carnitine palmityl transferase (CPT) 1, CTP2, dysferlin, thymidine phosphorylase; SMN2 (SMNC), insulin-like growth factor, sarcoplasmic reticulum Ca 2+ ATPase (SERCA), SERCA2a, phoshpholambin, histone deacetylases (HDACs), periostin, B-type natriuretic peptide (BNP), pseudophorphorylated mutant of phospholamban (S16EPLN), or Po 1 yADP ribose polymerase-1 (PARP-1).
18. The method according to claim 16 , wherein said transgene encodes Factor VIII or Factor IX.
19. The method according to claim 16 , wherein said transgene encodes a myostatin inhibitor, insulin, glucagon, growth hormone (GH), parathyroid hormone (PTH), growth hormone releasing factor (GRF), follicle stimulating hormone (FSH), luteinizing hormone (LH), human chorionic gonadotropin (hCG), vascular endothelial growth factor (VEGF), platelet derived growth factor (PDGF), angiopoietins, angiostatin, granulocyte colony stimulating factor (GCSF), erythropoietin (EPO), connective tissue growth factor (CTGF), basic fibroblast growth factor (bFGF), acidic fibroblast growth factor (aFGF), epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin growth factors I and II (IGF-I and IGF-II), TGFα, an activin, an inhibin, a bone morphogenic protein (BMP), a heregluin/neuregulin/ARIA/neu differentiation factor (NDF) growth factor, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin NT-3, neurotrophin NT-4/5, ciliary neurotrophic factor (CNTF), glial cell line derived neurotrophic factor (GDNF), neurturin, agrin, a semaphorin a collapsin, netrin-1, netrin-2, hepatocyte growth factor (HGF), an ephrin, noggin, sonic hedgehog or tyrosine hydroxylase.
20. The method according to claim 16 , wherein said transgene encodes FOS, CYR 61, IL-6, NR4A1, DUSP1, SLC2A3, MYC, IL8, IL-1beta, amphiregulin, PPP1R3C; a matrix metalloproteinase, Bc1-2, Folbpl, A63V, K70T, E180G alpha-tropomysin (Tm) mutations, FXR (farnesoid X receptor/bile acid receptor), a protein kinase C (PKC) serine/threonine kinase, parvalbumin (Parv), Gem, Adenylyl cyclase type VI (AC(VI)), human fibroblast growth factor 4 (Ad5FGF4)-adenoviral vector encoding HFGF4), cardiotrophin-1 (CT-1), Hsp90, troponin I and its isoforms, mutants, and chimeras, angiotensin II Type 2 Receptor (AT2R), S100A1, or human heme oxygenase-1.