This is a proposal that uses novel, systemically deliverable, small inhibitory peptides to determine whether combined targeting of neuron-intrinsic and extracellular inhibitory factors markedly improves retinal ganglion cell (RGC) axon regeneration and survival after optic nerve injury (ONI). Severed optic axons fail to regenerate and ONI leads to life-long visual loss in patients. In addition to apoptotic RGC death following axotomy, both a reduced intrinsic growth capacity and an inhibitory molecular environment contribute to failure of mature CNS neurons to regenerate their axons. Studies using conditional knockout (KO) mice suggest that the tumor suppressor gene PTEN is one neuron-intrinsic factor that critically restricts the regenerative capacity of adult RGCs. Deletion of PTEN enhanced RGC axon growth and survival after ONI. Chondroitin sulfate proteoglycans (CSPGs) generated in glial scars are extrinsic factors that strongly suppress axon extension. Recently, we and other labs identified LAR and PTP? phosphatases as receptors that mediate CSPG inhibition. Deletion of either of them partially overcomes suppression by CSPGs and stimulates growth of injured CNS axons. Some PTPs, including LAR, can also activate caspases and induce cell apoptosis. Suppressing PTEN and CSPG signaling is very promising for promoting CNS axon regeneration, but transgenic deletion of PTEN, LAR or PTP? is not feasible for treating patients. We have designed small mimetic peptides to block functions of these inhibitory molecules and demonstrated their efficiency in promoting axon growth in vitro and in vivo. Because CSPG receptors appear to regulate neuronal functions via pathways different from PTEN signaling, we hypothesize that combining inhibition of PTEN and CSPG signaling promotes RGC axon regeneration and survival better than inhibiting either alone. To test this hypothesis, we will use small peptide inhibitors alone and in combinations, validating the peptide efficacy by comparison to results with KO mice available in our lab. We propose to address 3 Specific Aims by determining whether: 1) peptide blockade of PTEN promotes similar RGC axon regeneration and survival as transgenic deletion in KO mice; 2) peptide blockade of two CSPG receptors promotes greater RGC axon regeneration and survival than inhibiting one receptor; and 3) blocking both PTEN and CSPG signaling with peptides promotes greater RGC axon regeneration and survival than targeting either one alone. By simultaneously targeting neuron-intrinsic and environmental inhibitory factors with small blocking peptides, we attempt to promote axon regeneration and to reduce axotomy-induced RGC loss to greater degrees than by targeting either signal individually. Our novel strategy of administering small, systemically deliverable compounds post-injury may facilitate development of practical combinatorial therapy for optic nerve injury.

Public Health Relevance

By comparison to RGC axon regeneration and survival in knockout mice, we will study the efficacy of our novel peptides on suppressing function of PTEN and two CSPG receptors. By simultaneously targeting neuron-intrinsic and CNS environmental inhibitory factors, we aim to promote better RGC axon regeneration and survival in adult rodents with optic nerve injury than by targeting either one alone. The invention and use of systemically deliverable blocking peptides may advance our ability to treat optic nerve injury by promoting axon regeneration.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY024575-02
Application #
8889260
Study Section
Diseases and Pathophysiology of the Visual System Study Section (DPVS)
Program Officer
Chin, Hemin R
Project Start
2014-08-01
Project End
2018-06-30
Budget Start
2015-07-01
Budget End
2016-06-30
Support Year
2
Fiscal Year
2015
Total Cost
Indirect Cost
Name
Temple University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
057123192
City
Philadelphia
State
PA
Country
United States
Zip Code
19122
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Sainath, Rajiv; Armijo-Weingart, Lorena; Ketscheck, Andrea et al. (2017) Chondroitin sulfate proteoglycans negatively regulate the positioning of mitochondria and endoplasmic reticulum to distal axons. Dev Neurobiol 77:1351-1370
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Ohtake, Yosuke; Wong, Daniella; Abdul-Muneer, P M et al. (2016) Two PTP receptors mediate CSPG inhibition by convergent and divergent signaling pathways in neurons. Sci Rep 6:37152
Feng, Dechun; Dai, Shen; Liu, Fengming et al. (2016) Cre-inducible human CD59 mediates rapid cell ablation after intermedilysin administration. J Clin Invest 126:2321-33
Ohtake, Yosuke; Li, Shuxin (2015) Molecular mechanisms of scar-sourced axon growth inhibitors. Brain Res 1619:22-35
Xu, Bin; Park, Dongsun; Ohtake, Yosuke et al. (2015) Role of CSPG receptor LAR phosphatase in restricting axon regeneration after CNS injury. Neurobiol Dis 73:36-48
Lang, Bradley T; Cregg, Jared M; DePaul, Marc A et al. (2015) Modulation of the proteoglycan receptor PTP? promotes recovery after spinal cord injury. Nature 518:404-8

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