Mechanisms of Microglia induced Retinal Degeneration

  • Xuan Du

Student thesis: Doctoral ThesisDoctor of Philosophy

Abstract

Retinal neurodegenerative diseases including age-related macular degeneration(AMD), retinitis pigmentosa (RP), diabetic retinopathy (DR) and glaucoma arecharacterized by irreversible vision loss and currently have very limited therapies.Evidence has suggested the immune profile and inflammatory etiology in patientswith retinal neurodegenerative diseases. Microglial activation is known to play animportant role in neuroinflammation and neurodegeneration. However, there is adebate on whether microglial activation is the primary cause or a consequence ofretinal neurodegeneration. It is also unclear why and how active microglia damageneurons since they are the main innate immune cells and their physiological role is toprotect the neuroretina.

Microglial activation is regulated by neurons. The chemokine CX3C motif receptor 1 (CX3CR1) is expressed exclusively by microglia in the central nerves system (CNS) and the retina; whereas the ligand, chemokine C-X3-C motif ligand 1 (CX3CL1) is expressed by neurons. The interaction between CX3CL1 and CX3CR1 is an important mechanism to avoid uncontrolled microglial activation. Microglia can be activated by inflammatory cytokines through the JAK/STAT pathway of cytokine receptors. Suppressor of cytokine signalling 3 (SOCS3) negatively regulates the JAK/STAT pathway. In this study, a mouse strain with the deletion of two inhibitory genes (CX3CR1 and SOCS3) in myeloid cells was created (named LysMCreSOCS3fl/flCX3CR1GFP/GFP, abbreviated as DKO in this thesis). The mice agedependently developed microglial activation and retinal neuronal degeneration.

Clinical examinations in young (3-5 months) and aged (10-12 months) wild-type(WT) control, LysMCre-SOCS3fl/fl, CX3CR1GFP/GFP and DKO mice showed that cluster of GFP signals and fundus abnormalities in aged DKO retina but not inretinas from other ages or genotypes. Immunohistochemistry showed severalalterations in aged DKO retina: 1) Uncontrolled microglial activation supported byincreased microglial cell number in all layers, altered distribution and lectinexpression; 2) Perturbed multiple retinal neurons including photoreceptors andbipolar cells and 3) Distorted RPE morphology (Chapter 3). 
To understand the mechanisms of active microglia-mediated retinal neuronal andRPE damage in the DKO mice, I investigated in Chapter 4 the phenotype andfunction of microglia from SOCS3fl/fl and DKO mice. The DKO microglia werefound to have following properties: 1) Enhanced phagocytic activity; 2) ElevatedmRNA expression of pro-inflammatory cytokines such as TNFα; 3) Upregulatedcytokine secretion in basal condition and in response to diverse stimuli and 4) DKOmicroglia aggravated the mortality rate of cone photoreceptors in retinal organoidculture. These data revealed that DKO microglia possessed stronger phagocyticpotential and cytotoxic ability, exacerbating photoreceptor death after differentstimuli.

To prove the retinal neuronal degeneration is primarily due to uncontrolledmicroglial activation, I used minocycline to control microglia activation in ourLysMCre-SOCS3fl/flCX3CR1GFP/GFP mice at the age of 5 months (before the onset ofretinal neuronal degeneration) for 3 consecutive months in Chapter 5. The treatmentsignificantly supressed microglial activation and prevented retinal neurodegeneration.Our data suggest that dysregulated microglial activation (e.g., due to genetic orepigenetic alteration) can be the primary cause of retinal neuronal degeneration. Ourstudy also suggests that active microglia may damage neurons by releasing cytotoxic cytokines such as TNFα, and CCL2. The knowledge is important for furtherdevelopment of strategies to control neuroinflammation and retinal degeneration.
Date of AwardJul 2020
Original languageEnglish
Awarding Institution
  • Queen's University Belfast
SponsorsQueen's University & China Scholarship Council
SupervisorMei Chen (Supervisor) & Heping Xu (Supervisor)

Cite this

Mechanisms of Microglia induced Retinal Degeneration
Du, X. (Author). Jul 2020

Student thesis: Doctoral ThesisDoctor of Philosophy