Abstract
Purpose :
The multipotent progenitor monocyte population in PBMCs have been used to generate cell which can be reconditioned called Reconditioned Monocytes (RM).These RMs have been sourced to differentiate in Retinal Neuron like Cells (RNLCs) in our lab. The RNLCs have shown to be a viable alternate to conventional cell-based therapy. Therefore, we aimed to analyse the potential of patient derived RNLCs for in vitro disease modelling for Retinitis Pigmentosa and a mutation specific in vitro model.
Methods :
Differentiating RP patient derived PBMCs into RNLCs was established by the two- step approach. Wherein, the PBMCs were isolated by density centrifugation method. The PBMC layer was collected and the adherent monocyte cells were supplemented with dedifferentiation media for 6 days. The reconditioned monocytes cells were then supplied with re-differentiation media until the cells attain the retinal phenotype at day 8.For the disease modelling study the cells were characterized by in vitro apoptosis profiling and compared with exome and clinical data.
Results :
Figure 1: Cumulative assessment of 40 RP patients against 30 healthy donors indicated that at D1 the monocytes were equally viable however apoptosis rates varied in the RP RLNCs with diverse patterns due to individual mutations’ appearance in the RLNCs and surmounting the effect. Thus, individual mutation specific patterns were studied.Figure 2: (A) RP1 displayed higher rate of apoptosis on day14 when compared to D6 an D10.(B) RP7 displayed higher rates of RNLC apoptosis on D6 while the RLNC were increasing viable at subsequent D10 and D14.
Conclusions :
Thus, the source retinal gene mutation manifests in the peripheral monocyte derived RLNCs and shows patterned apoptosis due the directional manifestation of the developmental genes. In vitro study of patient specific behaviour of retina towards degeneration can help understand the effect of any drug or cell-based therapy and mimic the patient condition to clinically characterize the rate of degeneration in a step towards targeted personalized medicine.
This is a 2021 ARVO Annual Meeting abstract.