Spinal cord injury (SCI) in humans leads to permanent paralysis because injured nerve fibers (axons) do not regenerate. One reason for this is the secretion of chondroitin sulfate proteoglycans (CSPGs) by cells near the injury. CSPGs bind to receptors of the protein tyrosine phosphatase (RPTP) family, PTP? and LAR, and inhibit axon growth. Regeneration after SCI is more successful in lampreys, but some identified reticulospinal (RS) neurons are bad regenerators, and when their axons are injured, these undergo very delayed programmed cell death (apoptosis). We will determine whether, acting through PTP? and/or LAR, CSPGs both inhibit true axon regeneration (as opposed to sprouting by uninjured axons) and trigger retrograde apoptosis after SCI. We will test whether digesting CSPGs with chondroitinase (ChABC) increases survival of spinal-projecting neurons and/or regeneration of their axons, and whether adding extrinsic CSPGs has the opposite effects. Evidence in vitro suggests that CSPGs, acting through LAR activate the small GTPase RhoA and inactivate Akt. Both signals have downstream effects that could inhibit axon growth. By blocking RhoA synthesis or activation and observing the effect on Akt activity, we will determine whether the effect on Akt is downstream of RhoA, or whether the two pathways are triggered independently, perhaps by different RPTPs. RhoA synthesis will be inhibited with morpholino antisense oligonucleotides (MOs) delivered to RS neurons retrogradely from the injury site. Activation of RhoA will be blocked with C3 transferase. The effect on apoptosis markers and axon regeneration will be determined. Synthesis of the RPTPs also will be inhibited with MOs, to determine which receptor mediates which of the negative effects of CSPGs. To test whether effects in lampreys also apply to mammalian neurons, we will perform parallel experiments on postnatal and adult mammalian primary neuronal cultures. We also will test in a mouse optic nerve crush model, whether genetic knockdown or pharmacological inhibition of RPTPs reduces retrograde neuronal death and enhances axonal regeneration in retinal ganglion cells in vivo. Understanding the intracellular pathways that mediate the inhibitory effects of CSPGs on cell survival and axon regeneration could lead to development of therapies for human SCI.

Public Health Relevance

Spinal cord injury (SCI) in humans leads to permanent paralysis because injured nerve fibers (axons) do not regenerate. In lampreys, axons regenerate, but some nerve cells are good regenerators and some bad. The bad regenerators also undergo very delayed programmed cell death after their axon is severed. Determining how the mechanisms for blocking regeneration relate to the mechanisms of cell death may help us to develop therapies for human SCI that both save nerve cells from dying and promote regeneration of their axons.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS092876-04
Application #
9670851
Study Section
Clinical Neuroplasticity and Neurotransmitters Study Section (CNNT)
Program Officer
Bambrick, Linda Louise
Project Start
2016-04-01
Project End
2021-03-31
Budget Start
2019-04-01
Budget End
2020-03-31
Support Year
4
Fiscal Year
2019
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
Zhang, Guixin; Hu, Jianli; Rodemer, William et al. (2018) RhoA activation in axotomy-induced neuronal death. Exp Neurol 306:76-91
Hu, Jianli; Zhang, Guixin; Rodemer, William et al. (2017) The role of RhoA in retrograde neuronal death and axon regeneration after spinal cord injury. Neurobiol Dis 98:25-35
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
Zhang, Guixin; Hu, Jianli; Li, Shuxin et al. (2014) Selective expression of CSPG receptors PTP? and LAR in poorly regenerating reticulospinal neurons of lamprey. J Comp Neurol 522:2209-29