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INFLAMMATION-DERIVED GROWTH FACTORS Optic Nerve Regeneration Is Becoming a Reality

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INFLAMMATION-DERIVED GROWTH FACTORS Optic Nerve Regeneration Is Becoming a Reality INFLAMMATION-DERIVED GROWTH FACTORS Optic nerve regeneration is becoming a reality. BY YUQIN YIN, MD, PHD; AND LARRY I. BENOWITZ, PHD ptic nerve trauma, ischemia, and or RAGs) in a pattern similar to that seen mannose, which is abundant in the vit- certain degenerative eye diseases during peripheral nerve regeneration.3 reous and cerebrospinal fluid, stimulates can lead to permanent vision Genetic deletion of two receptors appreciable axon growth from goldfish Oloss due to the inability of retinal that are expressed by inflammatory cells, RGCs and moderate outgrowth from rat ganglion cells (RGCs) to regenerate the Toll-like receptor 2 (TLR2) and dectin-1, RGCs. These effects require elevation of axons that convey visual information eliminates the proregenerative effects of cAMP and are strongly augmented by from the eye to the brain and the sub- zymosan, despite not altering the general a protein secreted by activated macro- sequent death of RGCs. Over the past profile of infiltrative cells.4 β-glucan is a phages.2,8 Using column chromatogra- 20 years, considerable progress has been component of zymosan that stimulates phy, mass spectrometry, and bioassays, made in defining factors that promote cells of the innate immune system via we identified the 11 kDa Ca2+-binding or suppress axon regeneration and RGC dectin-1, and curdlan, a particulate form protein oncomodulin (Ocm) as a major survival after optic nerve injury. of β-glucan, mimics the effects of zymo- growth-promoting factor associated This article describes research from san on regeneration.4 with inflammation.9 our lab that has identified trophic fac- Combining intraocular inflammation Ocm is secreted by infiltrative neutro- tors derived from inflammatory cells with elevation of cAMP and PTEN dele- phils and macrophages, and it accumu- that promote appreciable levels of tion in RGCs and other cells infected lates in the neural retina 12 to 24 hours optic nerve regeneration. These find- with an adeno-associated virus serotype after induction of intraocular inflamma- ings provide the basis for a potentially 2 (AAV2) expressing an anti-PTEN short tion, binding to a high-affinity receptor 5,9,10 viable gene therapy–based approach hairpin RNA (shRNA) has strongly syn- on RGCs (Kd ~ 28 nM). Elevation of that might in the future help enable ergistic effects. These include increasing cAMP alone induces only modest axon damaged retinal axons to grow back to axon regeneration 10-fold compared regeneration11 but is required for Ocm the brain and restore vision to patients with any of the treatments alone and and other trophic factors to bind to with optic nerve damage. enabling some axons to reach the their cognate receptors on RGCs.5,9,12,13 optic chiasm by 6 weeks5 and to rein- Delivery of Ocm and a cAMP analog INTRAOCULAR INFLAMMATION nervate subcortical visual nuclei by 10 via slow-release polymer beads mimics INDUCES OPTIC NERVE REGENERATION to 12 weeks.6 These regenerating axons the proregenerative effects of zymosan; We discovered that an unintentional become myelinated, although the pro- conversely, blocking the effects of Ocm injury to the lens induces considerable cess proceeds slowly.7 The brain target with either a neutralizing antibody axon regeneration and that this effect reinnervation leads to a limited recovery or a blocking peptide strongly sup- could be mimicked by inducing intraoc- of simple visual reflexes such as the presses the effects of zymosan.9,10,14 ular inflammation with zymosan, a frag- optomotor response.6 Regeneration is also diminished by ment of the yeast cell wall.1,2 This regen- These findings raise the question of immune-depletion of neutrophils, eration was found to be associated with whether the positive factors associated implying that these first responders of a change in RGCs’ intrinsic growth state, with inflammation can be identified the innate immune system mediate as evidenced by a massive upregulation to promote regeneration in a clinically most of the effects of inflammation on of GAP-43, SPRR1A, and other growth- useful way. optic nerve regeneration.10 associated proteins (GAPs; also called Oncomodulin. Our earlier work Ocm has also been reported to syn- regeneration-associated gene products, showed that the carbohydrate ergistically promote axon outgrowth NOVEMBER/DECEMBER 2019 | GLAUCOMA TODAY 39 s BEYOND IOP Figure. An example of a combinatorial gene therapy treatment leading to full-length optic nerve regeneration. CTB-positive axons are visualized in a longitudinal section through the optic nerve (14 µm thick) 6 to 8 weeks after nerve crush followed by intraocular injection of an adeno-associated virus expressing an shRNA to knock down expression of the PTEN gene (AAV2-shPTEN), a shH10-oncomodulin virus, AAV2-c/a-adenylate cyclase, and the chemokine stromal cell-derived factor 1 (SDF1). Asterisk: nerve injury site; scale bar: 150 µm. in RGCs when combined with a small inflammation doubles the number of trophic factors.38 One reason for the low interfering RNA against the Nogo-66 axons that regenerate the full length efficacy of rCNTF is that SOCS3, a repres- receptor15 and to contribute to the of the optic nerve and increases the sor of the Jak-STAT signaling pathway, conditioning lesion effect in the peripher- number of axons that extend through increases postnatally and increases even al nervous system. This, in turn, enables the optic chiasm and the optic tract further after optic nerve injury.3,32,39 injured dorsal root ganglion (DRG) and into the dorsal lateral geniculate Accordingly, deletion of SOCS3 ampli- neurons to extend axons following the nucleus.23 fies the effects of rCNTF.33 In addition, accumulation of infiltrative cells into SDF1 exerts its effects by activating mature RGCs do not express appre- peripheral nerves and DRGs.16 phosphatidylinositol 3-kinase (PI3K) ciable CNTFRa, the specific receptor SDF1. A second growth factor asso- signaling, elevating intracellular cAMP, subunit for CNTF (unpublished data). ciated with intraocular inflammation and antagonizing the axon repellant Elsewhere in the nervous system, this is the chemokine stromal cell-derived effects of slit/robo.23,25,26 Thus, Ocm subunit can be released from one factor 1 (SDF1; also called CXCL12). and SDF1 are two of the major prore- type of cell and become anchored to SDF1 acts through the receptor CXCR4, generative constituents of intraocular another cell type via a glycosylphospha- which is expressed in neurons, inflam- inflammation, and together they may tidylinositol linkage to form part of a matory cells, and other cell types17-19 be useful in promoting optic nerve tripartite receptor complex with LIFRβ as well as through CXCR7.20 SDF1 repair clinically. and glycoprotein 130 (gp130).40 has a wide range of effects on central CNTF. Ciliary neurotrophic factor CNTFRa is heavily expressed on nervous system development and (CNTF) has also been proposed to medi- astrocytes and inflammatory cells, and, hematopoiesis.21,22 It is highly expressed ate the effects of intraocular inflamma- as noted above, the effects of CNTF in infiltrative macrophages and acts tion on axon regeneration.27 Although gene therapy on optic nerve regenera- synergistically with Ocm to induce CNTF and other cardiotrophin family tion are mediated by factors secreted optic nerve regeneration.23 Deletion of chemokines become elevated in the by these cells. Nonetheless, CNTF and SDF1 in myeloid cells, using CXCL12fl/fl- eye after intraocular inflammation,10,28 related trophic factors appear to play LysMCre-/+ mice, or deletion of its recep- at physiologic concentrations recom- an important role in the visual system, tor CXCR4 in RGCs, using CXCR4fl/fl binant CNTF (rCNTF) alone has little as double deletion of CNTF and leuke- mice injected intraocularly with AAV2- axon-promoting effect on RGCs in cell mia inhibitory factor (LIF) accelerates Cre virus, diminished inflammation- culture9,14,29 and weak or no effects on RGC death after optic nerve injury and induced optic nerve regeneration by optic nerve regeneration in vivo.1,30-34 prevents regeneration.28 approximately one-third and fully elim- In the paradigm in which a segment of Other growth factors. Several other inated inflammation on RGC survival.23 peripheral nerve is grafted onto the cut growth factors have been reported to Blockade of both Ocm and SDF1 end of the optic nerve, high concentra- stimulate RGC survival but limited optic decreased inflammation-induced tions of rCNTF augmented axon regen- nerve regeneration; whether these fac- regeneration by 70% to 80%.23 In eration,35 but these effects were due to tors act directly on RGCs is generally gain-of-function experiments, although the chemotactic effects of CNTF on not known. These factors include, but SDF1 alone has only modest effects on macrophages.36,37 are not limited to, fibroblast growth regeneration,24,25 SDF1 combined with In contrast, unlike rCNTF, AAV2- factor-2 (FGF2),41 BDNF,31,42-49 GDNF,50,51 Ocm and cAMP mimics most of the mediated CNTF delivery induces con- and insulin-like growth factor 1 (IGF1)52- proregenerative effects of intraocular siderable axon regeneration through 55 combined with osteopontin.56 inflammation.23 The level of SDF1 asso- the optic nerve. However, we recently Gene therapy and full-length optic ciated with intraocular inflammation showed that this effect is due to the nerve regeneration. Combinatorial appears to be below optimal levels, as infiltration of inflammatory cells into the treatment of inflammation-derived adding exogenous SDF1 to intraocular eye that express Ocm, SDF1, and other growth factors via gene therapy 40 GLAUCOMA TODAY | NOVEMBER/DECEMBER 2019 BEYOND IOP s Acad Sci U S A. 2012;109(23):9149-9154. by cAMP analogue: impact on regeneration in injured retina. Mol Cell Neurosci. has potential clinical significance. 7. Marin MA, de Lima S, Gilbert HY, Giger RJ, Benowitz L, Rasband MN. Reassembly 2009;41(3):313-324.
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