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Targeting chondroitinase ABC to axons enhances the ability of chondroitinase to promote neurite outgrowth and sprouting


Autoři: Priscilla Day aff001;  Nuno Alves aff002;  Esther Daniell aff001;  Debayan Dasgupta aff001;  Rosalie Ogborne aff001;  Ashley Steeper aff001;  Mansoor Raza aff001;  Clare Ellis aff001;  James Fawcett aff002;  Roger Keynes aff001;  Elizabeth Muir aff001
Působiště autorů: Department of Physiology Development and Neuroscience, University of Cambridge, Cambridge, United Kingdom aff001;  John Van Geest Centre for Brain Repair, University of Cambridge, Cambridge, United Kingdom aff002
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0221851

Souhrn

Background

There is currently no effective treatment for promoting regeneration of injured nerves in patients who have sustained injury to the central nervous system such as spinal cord injury. Chondroitinase ABC is an enzyme, which promotes neurite outgrowth and regeneration. It has shown considerable promise as a therapy for these conditions. The aim of the study is to determine if targeting chondroitinase ABC expression to the neuronal axon can further enhance its ability to promote axon outgrowth. Long-distance axon regeneration has not yet been achieved, and would be a significant step in attaining functional recovery following spinal cord injury.

Methodology/Principal findings

To investigate this, neuronal cultures were transfected with constructs encoding axon-targeted chondroitinase, non-targeted chondroitinase or GFP, and the effects on neuron outgrowth and sprouting determined on substrates either permissive or inhibitory to neuron regeneration. The mechanisms underlying the observed effects were also explored. Targeting chondroitinase to the neuronal axon markedly enhances its ability to promote neurite outgrowth. The increase in neurite length is associated with an upregulation of β-integrin staining at the axonal cell surface. Staining for phosphofocal adhesion kinase, is also increased, indicating that the β-integrins are in an activated state. Expression of chondroitinase within the neurons also resulted in a decrease in expression of PTEN and RhoA, molecules which present a block to neurite outgrowth, thus identifying two of the pathways by which ChABC promotes neurite outgrowth.

Conclusions / Significance

The novel finding that targeting ChABC to the axon significantly enhances its ability to promote neurite extension, suggests that this may be an effective way of promoting long-distance axon regeneration following spinal cord injury. It could also potentially improve its efficacy in the treatment of other pathologies, where it has been shown to promote recovery, such as myocardial infarction, stroke and Parkinson’s disease.

Klíčová slova:

Axons – Cell staining – Integrins – Neurites – Neuronal differentiation – Neurons – Spinal cord injury – Transfection


Zdroje

1. Snow DM, Lemmon V, Carrino DA, Caplan AL, Silver J. Sulfated proreoglycans in astroglial barriers inhibit neurite outgrowth in vitro. Exp Neurol (1999); 109:111–130.

2. Silver J & Miller JH. Regeneration beyond the glial scar. Nat Rev Neurosci (2004); 5:146–156. doi: 10.1038/nrn1326 14735117

3. Morgenstern DA, Asher RA, Fawcett JW. Chondroitin sulphate proteoglycans in the CNS injury response, Prog Brain Res.(2002); 137:313–332. doi: 10.1016/s0079-6123(02)37024-9 12440375

4. Bradbury EJ, Moon LD, Popat RJ, King VR, Bennet GS, Patel PN, et al. Chondroitinase ABC promotes functional recovery after spinal cord injury. Nature (2002); 416: 636–40. doi: 10.1038/416636a 11948352

5. Zhao RR, Muir EM, Alves JN, Rickman H, Allan AY, Kwok JC, et al. Lentiviral vectors express Chondroitinase ABC in cortical projections and promote sprouting of injured corticospinal axons. J Neurosci Methods (2011); 201:228–238. doi: 10.1016/j.jneumeth.2011.08.003 21855577

6. Massey JM, Hubscher CH, Wagoner MR, Decker JA, Amps J, Silver J, et al. Chondroitinase ABC digestion of the perineuronal net promotes functional collateral sprouting in the cuneate nucleus after cervical spinal cord injury. J Neurosci. (2006); 26: 4406–4414. doi: 10.1523/JNEUROSCI.5467-05.2006 16624960

7. Mondello SE, Jefferson SC, Tester NJ, Howland DR. Impact of treatment duration and lesion size on effectiveness of ChABC treatment post-spinal cord injury. Exp Neurol. (2015); 267: 64–77. doi: 10.1016/j.expneurol.2015.02.028 25725355

8. Lee HJ, Bian S, Jakovcevski I, Wu B, Irintchev A, Schachner M. Delayed applications of L1 and ChABC promote recovery after spinal cord injury. J Neurotrauma (2012); 29: 1850–1863. doi: 10.1089/neu.2011.2290 22497349

9. Garcia-Alias G, Barkhuysen S, Buckle M, Fawcett JW. Chondroitinase ABC treatment opens a window of opportunity for task-specific rehabilitation. Nat Neurosci. (2009); 12:1145–51. doi: 10.1038/nn.2377 19668200

10. Sivak WN, White JD, Biley JM, Tien LW, Liao HT, Kaplan DC et al. Delivery of ChondroitinaseABC and glial cell line-derived neurotrophic factor from silk fibroin conduits enhances peripheral nerve regeneration. J Tissue Eng Regen Med. (2014); doi: 10.1002/term.1970 25424415

11. Gardner RT, Habecker BA. Infarct-derived chondroitin sulfate proteoglycans prevent reinnervation after cardiac ischemia-reperfusion injury. J Neurosci. (2013); 33: 7175–7183. doi: 10.1523/JNEUROSCI.5866-12.2013 23616527

12. Chen XR, Liao SJ, Ye LX, after stroke in hypertensive rats. Gong Q, Ding Q, Zeng JS, et al. Neuroprotective effect of Chondroitinase ABC on primary and secondary brain injury Brain Res. (2014); 1543: 324–333. doi: 10.1016/j.brainres.2013.12.002 24326094

13. Gheardini L, Gennaro M, Pizzorusso T. Perilesional treatment with ChABC ABC and motor training promote functional recovery after stroke in rats. Cereb Cortex (2015); 1: 202–212.

14. Muir EM, Fyfe I, Gardiner S, Li L, Warren P, Fawcett J, et al. Modification of N-glycosylation sites allows secretion of bacterial chondroitinase ABC from mammalian cells. J Biotechnol. (2010);145: 103–110. doi: 10.1016/j.jbiotec.2009.11.002 19900493

15. Bartus K, James ND, Didangelos A, Bosch KD, Verhaagen J, Yáñez-Muñoz RJ, et al. "Large-scale chondroitin sulfate proteoglycan digestion with Chondroitinase ABC gene therapy leads to reduced pathology and modulates macrophage phenotype following spinal cord contusion injury." J Neurosci. (2014); 34: 4822–4836. doi: 10.1523/JNEUROSCI.4369-13.2014 24695702

16. Agholme L, Lindstrom T, Kagedal K, Marcussion J, Hallbeck M. An in vitro model for neuroscience: differentiation of SH-SY-5Y cells into cells with morphological and biochemical characteristics of mature neurons. J. Alzheimer’s dis. (2010); 20: 1069–1082.

17. Satpute-Krishnan P, DeGiorgis JA, Conley MP, Jang M, Bearer EL. A peptide zipcode sufficient for anterograde transport within amyloid precursor protein. Proc Natl Acad Sci. (2006);103:16532–16537. doi: 10.1073/pnas.0607527103 17062754

18. Wang H, Katagiri Y, McCann TE, Unsworth E, Goldsmith P, Yu ZX, et al. Chondroitin-4-sulfate negatively regulates axonal guidance and growth. J Cell Sci. (2008); 121:3083–3091. doi: 10.1242/jcs.032649 18768934

19. Caceres A, Banker GA, Binder L. Immunocytochemical localization of tubulin and microtubule-associated protein 2 during the development of hippocampal neurons in culture. J Neurosci. (1986); 6:714–722. 3514816

20. Katsetos CD, Hermann MM, Mork SJ. Class III β-tubulin in human development and cancer. Cell Motil Cytoskeleton (2003); 55:77–96. doi: 10.1002/cm.10116 12740870

21. Richardson A and Parsons T. A mechanism for regulation of the adhesion-associated protein tyrosine kinase pp125 FAK. Nature (1996); 380: 538–540. doi: 10.1038/380538a0 8606775

22. Gavet O and Pines J. Progressive activation of cyclin B1-cdk1 coordinates entry to mitosis. Dev Cell (2010);18:533–543. doi: 10.1016/j.devcel.2010.02.013 20412769

23. Holm M. (1979). A simple sequentially rejective multiple test procedure. Scand J Statistics 6:65–70

24. Sackett S, Tremmel D, Ma F. Extracellular matrix scaffold and hydrogel derived from decelluarized and delipidized human pancreas. Sci rep (2018); doi: 10.1038/s41598-018-28857-1

25. Rosivatz E, Matthews JG, McDonald NQ, Mulet X, Ho KK, Lossi N, et al. A small molecule inhibitor for phosphatase and tensin homologue deleted on chromosome 10 (PTEN). ACS Chem Biol. (2006); 1: 780–790. doi: 10.1021/cb600352f 17240976

26. Kopp MA, Liebscher T, Niedeggan A, Laufen S, Brommer B, Jungehulsing GJ, et al. Small-molecule-induced Rho-inhibition NSAIDS after spinal cord injury. Cell Tissue Res. (2012); 348: 119–132. doi: 10.1007/s00441-012-1360-5

27. Babetto E, Beirowski B, Janeckova L, Brown R, Gilley J, Thomson D, et al. Targeting NMNAT1 to axons and synapses transforms its neuroprotective potency in vivo. J Neurosci. (2010); 30: 13291–13304. doi: 10.1523/JNEUROSCI.1189-10.2010 20926655

28. Muir E, Raza M, Ellis C, Burnside E, Love F, Heller S. et al. Trafficking and processing of bacterial proteins by mammalian cells: Insights from chondroitinase ABC. (2017); doi: 10.1371/journal.pone.0186759 29121057

29. Lander C, Zhang H, Hockfield S. Neurons produce a neuronal cell surface-associated chondroitin sulfate proteoglycan. J Neurosci. (1998); 18: 174–183. 9412498

30. Orlando C, Ster J, Gerber U, Fawcett JW, Rainetreau O. Perisynaptic chondroitin sulfate proteoglycans restrict structural plasticity in an integrin-dependent manner. J Neurosci. (2012); 32: 18009–18017. doi: 10.1523/JNEUROSCI.2406-12.2012 23238717

31. Tan CL, Kwok JC, Patani R, Ffench-Constant C, Chandran S, Fawcett JF. Integrin activation promotes axon growth on inhibitory chondroitin sulphate proteoglycans by enhancing integrin signalling. J Neurosci. (2011); 31: 6289–6295. doi: 10.1523/JNEUROSCI.0008-11.2011 21525268

32. Andrews MR, Czvitovich S, Dassie E, Vogelaar CF, Faissner A, Blits B, et al. Alpha9 integrin promotes neurite outgrowth on tenasin-C and enhances axon regeneration. J Neurosci. (2009); 29: 5546–5557. doi: 10.1523/JNEUROSCI.0759-09.2009 19403822

33. Walker BA, Ji SJ, Jaffrey SR. Intra-axonal translation of RhoA promotes axon growth inhibition by CSPG. J Neurosci Methods (2012a); 201:228–238.

34. Walker BA, Hengst U, Kim HJ, Jeon NL, Schmidt EF, Heintz N, et al. Reprogramming axonal behaviour by axon-specific viral transduction. Gene Ther. (2012b); 19: 947–955.

35. Mak LH and Woscholski R. Targeting PTEN using small molecule inhibitors. Methods (2015);77–78: 63–68. doi: 10.1016/j.ymeth.2015.02.007 25747336

36. Perdigoto AL, Chaudhry N, Barnes GN, Filbin MT, Carter BD. A novel role for PTEN in inhibition of neurite outgrowth by myelin-associated glycoprotein in cortical neurons. Mol Cell Neurosci. (2011);46: 235–244. doi: 10.1016/j.mcn.2010.09.006 20869442

37. Liu K, Lu Y, Lee JK, Samara R, Willenberg R, Sears-Kraxberger I, et al. PTEN deletion enhances the regenerative ability of adult corticospinal neurons. Nat Neurosci. (2010);13:1075–1081. doi: 10.1038/nn.2603 20694004

38. Lee DH, Luo X, Yunger BJ, Bray E, Lee JK, Park KK. Mammalian target of rapamycin’s distinct roles and effectiveness in promoting compensatory axonal sprouting in the injured CNS. J Neurosci. (2014); 34:15347–15355. doi: 10.1523/JNEUROSCI.1935-14.2014 25392502

39. Danilov CA and Steward O. Conditional genetic deletion of PTEN after spinal cord injury enhances regenerative growth of CST axons and motor function recovery in mice. Exp Neurol. (2015); 26:147–160.


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