Brain protection using autologous bone marrow cell, metalloproteinase inhibitors, and metabolic treatment in cerebral ischemia

doi:10.1073/pnas.0611112104

Brain protection using autologous bone marrow cell, metalloproteinase inhibitors, and metabolic treatment in cerebral ischemia

*British Heart Foundation Glasgow Cardiovascular Research Centre, 126 University Place, Glasgow G12 8TA, United Kingdom;
^†Department of General Pathology, Excellence Center on Cardiovascular Diseases, and Research Center on Craniofacial Malformations–MRI, First School of Medicine, Second University of Naples, Naples 80138, Italy;
^‡Division of Anesthesiology and
^¶Department of Molecular Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095; and
^§Division of Hypertension, Mayo Clinic Foundation, Rochester, MN 55095

Contributed by Louis J. Ignarro, December 22, 2006 (received for review October 30, 2006)

Abstract

Despite advances in imaging, understanding the underlying pathways, and clinical translation of animal models of disease there remains an urgent need for therapies that reduce brain damage after stroke and promote functional recovery in patients. Blocking oxidant radicals, reducing matrix metalloproteinase-induced neuronal damage, and use of stem cell therapy have been proposed and tested individually in prior studies. Here we provide a comprehensive integrative management approach to reducing damage and promoting recovery by combining biological therapies targeting these areas. In a rat model of transient cerebral ischemia (middle cerebral artery occlusion) gene delivery vectors were used to overexpress tissue inhibitor of matrix metalloproteinase 1 and 2 (TIMP1 and TIMP2) 3 days before ischemia. After occlusion, autologous bone marrow cells alone or in combination with agents to improve NO bioavailability were administered intraarterially. When infarct size, BrdU incorporation, and motor function recovery were determined in the treatment groups the largest beneficial effect was seen in rats receiving the triple combined therapy, surpassing effects of single or double therapies. Our study highlights the utility of combined drug, gene, and cell therapy in the treatment of stroke.

Footnotes

^‖To whom correspondence may be addressed. E-mail: lignarro{at}mednet.ucla.edu or claunap{at}tin.it
Author contributions: L.J.I. and C.N. designed research; A.H.B., V.S., L.M.W., S.W.-I., F.d.N., L.O.L., A.C., A.L., C.S., and M.R. performed research; A.H.B. and L.M.W. contributed new reagents/analytic tools; A.H.B., V.S., L.M.W., S.W.-I., F.d.N., L.O.L., A.C., A.L., C.S., M.R., L.J.I., and C.N. analyzed data; and A.H.B., L.M.W., L.O.L., L.J.I., and C.N. wrote the paper.
Conflict of interest statement: L.J.I. helped develop and has a financial interest in a commercially available dietary supplement that contains some of the amino acids and antioxidants studied in this article.
Abbreviations:

BBB,

blood–brain barrier;

BMC,

bone marrow cell;

MCAO,

middle cerebral artery occlusion;

MMP,

matrix metalloproteinase;

TIMP,

tissue inhibitor of matrix metalloproteinase;

IBZ,

ischemic boundary zone;

TTC,

2,3,5-triphenyltetrazolium chloride;

GFAP,

glial fibrillary acidic protein;

VWF,

von Willebrand factor;

MAP-2,

microtubule-associated protein 2;

Neu-N,

neuronal nuclei;

NOx,

nitrite plus nitrate.