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@PHDTHESIS{Panten:295846,
author = {J. Panten$^*$},
title = {{C}auses and consequences of context-specific allelic
imbalance},
school = {Universität Heidelberg},
type = {Dissertation},
reportid = {DKFZ-2024-02663},
year = {2024},
note = {Dissertation, Universität Heidelberg, 2024},
abstract = {Gene expression has to be regulated in a cell type-specific
manner to ensure proper functionality of cell types and
tissues. In a diploid organism, the two alleles of a gene
can be regulated independently, causing differential
contribution to mRNA levels and thus to cellular function.
Allelic imbalance in gene expression has long been
recognized as a contributor to cellular phenotypes, however,
it is not well understood how and to which extent allelic
imbalance is shaped by the regulatory environment of
different cell types. Recent advances in single-cell
technologies provide the opportunity to profile gene
expression and its regulation in a cell type-specific manner
at scale, extending our understanding of genome function. In
this thesis, I performed a comprehensive analysis of
allele-specific expression (ASE) at single-cell resolution
in interspecific mouse hybrids. I first analysed the
differentiation-dependence of allelic imbalance caused by
strain-specific genetic effects during murine
spermatogenesis. This analysis shows that across cell types,
variation in ASE is extremely pervasive. Using an F1 trio
design, I further separated cis- and trans-contributions to
gene expression divergence and showed that cell
type-specific action of regulatory variants is mainly driven
by the interaction of cis-effects with the cellular
environment. Finally, I investigated the contribution of
dynamic genetic effects to cell type-specific
transcriptional evolution. Next, I focussed on ASE caused by
an epigenetic mechanism, namely X-chromosome inactivation
(XCI). In female humans and mice, XCI causes mosaic
haplotype-specific expression of X-linked genes, and escape
from XCI can lead to increased gene dosage compared to
males. Using single-cell genomics assays, I developed an
analysis approach to distinguish active and inactive
X-chromosomes in individual cells, which allowed me to
identify cell type-specific escape. I further showed that
T-cell expansion during ageing leads to globally impaired
silencing of the inactive X which is associated with an
exhaustion phenotype. These findings replicated on the level
of chromatin accessibility, demonstrating that variation in
escape is associated with an active chromatin state.
Collectively, I showed that escape can vary at the cell type
level and during organismal ageing. While these results show
that escape from XCI is plastic, they do not address how it
might be regulated in different cell types. In the final
chapter, I therefore explored whether the Xist long
non-coding RNA can regulate escapee expression. Using
allele-specific RNA-Seq data, I showed that increased
Xist-levels lead to almost complete silencing of escapees in
neural progenitor cells. Modelling of silencing trajectories
showed substantial variability among genes in both their
resistance to silencing as well as their reversibility,
suggesting that escape is genomically encoded. Finally, I
demonstrated that over-expression of Xist leads to escapee
silencing in early embryogenesis. These results provide a
potential mechanism that might drive variability in
expression from the inactive X. Taken together, this thesis
delineates to which extent allelic imbalance is driven by
cell typespecific regulatory environments and suggests
analysis approaches for allele-resolved single-cell data.
This provides the basis for a comprehensive survey of
allelic usage in vivo and the molecular mechanisms causing
its context-specificity.},
cin = {B260 / B270},
cid = {I:(DE-He78)B260-20160331 / I:(DE-He78)B270-20160331},
pnm = {312 - Funktionelle und strukturelle Genomforschung
(POF4-312)},
pid = {G:(DE-HGF)POF4-312},
typ = {PUB:(DE-HGF)11},
url = {https://inrepo02.dkfz.de/record/295846},
}