A major barrier to regeneration of CNS axons is the presence of growth-inhibitory proteins associated with myelin debris and the glial scar. Functional recovery after CNS injury requires that this inhibition be overcome. Recent studies suggest that changes in cAMP, along with increases in PKC, EGFR, and RhoA activities, are important aspects of inhibitory signaling. However, we still lack knowledge about the number/identity of inhibitory proteins associated with inhibition at injury sites, the detailed signaling mechanisms employed by inhibitory receptors, and the cell type-specific responses of damaged axons. Further, there are problems associated with current pharmacological strategies, including lack of specificity, uncertain toxicities, and the targeting of pathways with pleiotrophic functions. To overcome these difficulties, we have initiated a phenotype-based unbiased screen of a novel chemical compound library chosen for its favorable chemical properties rather than known biological function. The screen is based on the ability of compounds to increase neurite outgrowth from CNS neurons challenged with inhibitory myelin substrates. Initial results have produced 4 """"""""hit compounds"""""""" capable of strongly increasing neurite growth. Subsequent investigations indicate that the hit compounds a) act on different neuronal types, b) selectively overcome inhibition rather than promote growth, c) are highly potent, d) overcome inhibition in distinct assays relevant to injury, e) do not affect cAMP levels, PKC activity, or EGFR activation, f) alter microtubule dynamics, and g) promote regeneration in vivo. Because the compounds are potent and selective, and may act through novel mechanisms, they are exciting candidates for therapeutic development and for mechanistic studies of regeneration inhibition. The proposal is to 1) investigate the signaling mechanisms and protein targets of the 4 hit compounds, 2) examine the ability of 1 hit compound to promote regeneration after spinal cord injury or optic nerve crush in vivo, and 3) screen the full 4000 compound library on a novel inhibitory (proteoglycan) substrate. These experiments could provide key insights into regeneration inhibition, and pave the way for a novel approach to CNS injury.

Public Health Relevance

The proposed experiments will investigate the mechanisms of action of novel compounds promoting regeneration, and elucidate their ability to increase axonal regrowth after CNS injury.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS059866-03
Application #
7991788
Study Section
Special Emphasis Panel (ZRG1-MDCN-N (02))
Program Officer
Kleitman, Naomi
Project Start
2009-01-01
Project End
2013-12-31
Budget Start
2011-01-01
Budget End
2011-12-31
Support Year
3
Fiscal Year
2011
Total Cost
$327,994
Indirect Cost
Name
University of Miami School of Medicine
Department
Pharmacology
Type
Schools of Medicine
DUNS #
052780918
City
Coral Gables
State
FL
Country
United States
Zip Code
33146
Ruschel, Jörg; Hellal, Farida; Flynn, Kevin C et al. (2015) Axonal regeneration. Systemic administration of epothilone B promotes axon regeneration after spinal cord injury. Science 348:347-52
Lerch, Jessica K; Martínez-Ondaro, Yania R; Bixby, John L et al. (2014) cJun promotes CNS axon growth. Mol Cell Neurosci 59:97-105
Gatto, Graziana; Dudanova, Irina; Suetterlin, Philipp et al. (2013) Protein tyrosine phosphatase receptor type O inhibits trigeminal axon growth and branching by repressing TrkB and Ret signaling. J Neurosci 33:5399-410
Al-Ali, Hassan; Schürer, Stephan C; Lemmon, Vance P et al. (2013) Chemical interrogation of the neuronal kinome using a primary cell-based screening assay. ACS Chem Biol 8:1027-36
Blackmore, Murray G; Wang, Zimei; Lerch, Jessica K et al. (2012) Kruppel-like Factor 7 engineered for transcriptional activation promotes axon regeneration in the adult corticospinal tract. Proc Natl Acad Sci U S A 109:7517-22
Lerch, Jessica K; Kuo, Frank; Motti, Dario et al. (2012) Isoform diversity and regulation in peripheral and central neurons revealed through RNA-Seq. PLoS One 7:e30417
Nelersa, Claudiu M; Barreras, Henry; Runko, Erik et al. (2012) High-content analysis of proapoptotic EphA4 dependence receptor functions using small-molecule libraries. J Biomol Screen 17:785-95
Lerch, Jessica K; Bixby, John L; Lemmon, Vance P (2012) Isoform diversity and its importance for axon regeneration. Neuropathology 32:420-31
Motti, Dario; Bixby, John L; Lemmon, Vance P (2012) MicroRNAs and neuronal development. Semin Fetal Neonatal Med 17:347-52
Slepak, Tatiana I; Salay, Lindsey D; Lemmon, Vance P et al. (2012) Dyrk kinases regulate phosphorylation of doublecortin, cytoskeletal organization, and neuronal morphology. Cytoskeleton (Hoboken) 69:514-27

Showing the most recent 10 out of 20 publications