Abstract
Cell division refers to the process by which cells grow, replicate their genetic
material, and divide to form daughter cells. Major biological processes, namely
reproduction, development, wound healing and tissue regeneration, require cell division.
Cells switch between quiescence and proliferation states for maintaining tissue
homeostasis and regeneration. The cell division process is regulated by a wide range of
extracellular and intracellular cues like growth factors, stress, and reactive oxygen species
(ROS). The decision to exit or enter quiescence is dysregulated in proliferative and
degenerative diseases. Hence, understanding the molecular mechanisms that control the
reversible transition between quiescence and proliferation is crucial. In this thesis, we
study the regulatory network involved in this decision-making in normal and disease
(cancers and Alzheimer’s Disease) conditions and characterize the metabolic adaptation
of cancers using systems biology approach.
At the restriction point (R-point), cells become irreversibly committed to the
completion of the cell cycle independent of mitogen. The mechanism involving hyperphosphorylation of retinoblastoma (Rb) and activation of transcription factor E2F is
linked to the R-point passage. However, stress stimuli trigger exit from the cell cycle back
to the mitogen-sensitive quiescent state after Rb hyper-phosphorylation, but only until
APC/C-Cdh1 inactivation. In the work presented here, we developed a mathematical
model to investigate the reversible transition between quiescence and proliferation in
mammalian cells with respect to mitogen and stress signals. The model integrates the
current mechanistic knowledge and accounts for the recent experimental observations
with cells exiting quiescence and proliferating cells. We show that Cyclin E-Cdk2
couples Rb-E2F and APC/C-Cdh1 bistable switches and temporally segregates the Rpoint and the G1/S transition. A redox-dependent mutual antagonism between APC/CCdh1 and its inhibitor Emi1 makes the inactivation of APC/C-Cdh1 bistable. We show
that the levels of Cdk inhibitor (CKI) and mitogen control the reversible transition
between quiescence and proliferation. Further, we propose that shifting of the mitogeninduced transcriptional program to G2-phase in proliferating cells might result in an
intermediate Cdk2 activity at the mitotic exit and the immediate inactivation of APC/CCdh1. Our study builds a coherent framework and generates hypotheses that have been confirmed by experimental findings.
Proliferative diseases like cancer arise due to alterations in the regulation of the
cell cycle. An emerging hallmark of cancer is metabolic reprogramming, which presents
opportunities for cancer diagnosis and treatment based on metabolism. A comprehensive
metabolic network analysis of renal cell carcinoma (RCC) subtypes, including clear cell,
papillary, and chromophobe, was performed by integrating transcriptome data with the
human genome-scale metabolic model to understand the coordination of metabolic
pathways in cancer cells. We identified metabolic alterations of each subtype with respect
to tumor-adjacent normal samples and compared them to understand the differences
between subtypes. We found that genes of amino acid metabolism and redox homeostasis
are significantly altered in RCC subtypes. Chromophobe showed metabolic divergence
compared to other subtypes with upregulation of genes involved in glutamine anaplerosis
and aspartate biosynthesis. A difference in transcriptional regulation involving HIF1A is
observed between subtypes. We identified E2F1 and FOXM1 as other major
transcriptional activators of metabolic genes in RCC. These results highlight the crosstalk
between metabolism and cell division. Further, the co-expression pattern of metabolic
genes in each patient showed variations in metabolism within RCC subtypes. We also
found that co-expression modules of each subtype have tumor stage-specific behavior,
which may have clinical implications.
Intriguingly, cell cycle dysregulation triggers not only proliferative diseases such
as cancers but also drives degenerative diseases like Alzheimer's disease (AD). Aberrant
production and aggregation of amyloid beta oligomers (Aβ) into plaques is a frequent
feature of AD. However, therapeutic approaches targeting Aβ accumulation fail to reverse
or inhibit disease progression. The approved cholinesterase inhibitor drugs are also
mostly symptomatic treatments. During human brain development, the progenitor cells
differentiate into neurons and switch to a postmitotic, resting state. However, cell cycle
re-entry often precedes the loss of neurons. In this study, we developed mathematical
models of multiple routes leading to cell cycle re-entry in neurons that incorporate the
crosstalk between cell cycle, neuronal and apoptotic signaling mechanisms. We show that
the integration of multiple feedback loops influenc