advanced-stem-cell-2018-posters-abstracts

Advanced Stem Cell 2018

Journal of Stem Cell Biology and Transplantation

ISSN: 2575-7725

Page 43

December 03-04, 2018

Valencia, Spain

Edition of EuroSciCon Conference on

Advanced Stem Cell &

Regenerative Medicine

he active immunity in an immune compromised or radiation

victim can be restored by generating functionally competent

immune cells. Transcription factor like PU.1 plays a critical role

in the differentiation of hematopoietic stem cells (HSCs) into

immune cells. Another transcription factor GATA-1 (43 kDa)

interacts with PU.1, which promotes differentiation of HSCs into

erythroid lineages by blocking the binding of its coactivator c-Jun

with β3/β4 region of PU.1. We have established an antagonizing

mutation(Y244D)inβ3/β4domainbysite-directedmutagenesisof

PU.1, anticipating the enhancement of myelopoiesis if introduced

into cells. We further assessed if it can really antagonize GATA-

1 and promote myelopoiesis in cells upon transfection of native

and mutant PU.1 protein formulated into nanoparticle structure.

Approximately 30% of human bone-marrow CD34+ stem cells

were transformed into myeloid lineage by using these NPs loaded

with PLGA in vitro. In vivo studies by infusing primed CD34+ with

these NPs and along with cytokines were conducted in irradiated

nude mice. Results demonstrated higher survival rate (50-75%)

in mice compared to control mice with a reasonable number of

macrophages (43.11%) and neutrophils (50.44%) being observed

on the 21st day. Further high-throughput microarray analysis

in CD34+ cells whose selective differentiation was induced by

PU.1 (Y244D) mutant transfection was performed to explore

the myeloid specific gene-cluster responsible for myeloid

development. Gene expression analysis demonstrated that a ≥2.5

fold change were observed in several genes like PRTN3, TLR4,

EPAS1, FCER1A, NOD2, CBL, SMAD4, ANO6, TFRC, IL1R1 etc.

Studies on establishing the role/mechanistic basis of these genes

are presently under progress.

Harnessing PU.1 and GATA-1 axis for priming

human CD34+ stem cells towards myeloid

lineages

Madhuri Joshi

and

Gurudutta Gangenahalli

INMAS-DRDO, India

J Stem Cell Biol Transplant 2018, Volume 2

DOI: 10.21767/2575-7725-C1-003