Supervisor:  Dr David Connolly  

Department: Clinical Science and Services

Hypothesis

Microvesicles derived from hypoxic preconditioned Cardiosphere derived cells will have enhanced anti-apoptotic, reparative and pro-angiogenic activity measured by established in vitro functional assays.

Aims

Compare the efficacy of microvesicles (MVs) from hypoxic preconditioned (HP) cardiosphere derived cells (CDCs) with MVs from CDCs cultured at 21% 02 to mitigate injury responses using established in vitro models.

Background

Canine cardiomyopathy cause significant morbidity and mortality. Myocardial inflammation is the key to disease initiation and progression. Cardiosphere derived cells (CDCs) are used therapeutically but efficacy to date has been sub-optimal potentially because CDCs cause only transient benefit due to their rapid elimination from the myocardium. Stem cell therapy acts via paracrine mechanisms that can be exploited by isolating microvesicles (MVs) which can be repeatedly administered intravenously enabling prolonged therapeutic benefit. In a pivotal study systemic delivery of MVs from human CDCs in a mouse DCM model improved heart function and angiogenesis and decreased apoptosis and fibrosis. Therapeutic potential can be improved by strategies to enhance the anti-inflammatory, anti-apoptotic and proangiogenic activity of MVs derived from CDCs by hypoxic preconditioning (HP) driving upregulation of hypoxia inducible factor (HIF).

Project Timeline

2018/19 2019 2019
Task 1

Optimise culture of CDCs atvariable O2 concentrations (1-5%) to determine the cell growth rate and survival.

Perform HIF-1 analysis, identify, and measure HIF-1 regulated pathways.

Isolate quantify and store MVs for functional assays

Oct - Jan
Task 2

Optimise and perform functional assays:

Scratch assay to measure the efficacy of MVs for wound healing.

Apoptosis assay to measure the capacity of MVs to modify apoptosis pathways.

Angiogenesis assay to measure pro-angiogenic capacity of MVs.

Feb - July
Task 3 Write dissertation – prepare manuscript for publication August

This is a full-time (12 month) project commencing in October 2018.

The ECVIM clinical studies fund has been secured to cover consumable costs. The MRes student will be expected to meet the outstanding sum required, course fees and their living expenses.

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