ETec_Lecture4: Tissue dynamics
Tissue dynamics
The 3 dynamic states of tissues and the underlying cellular-fate processes.
• Tissue function (homeostasis – equilibrium)
o The normal steady-state function of tissue
▪ Some tissues produce cells as their main function
▪ Produced a secreted product
▪ Carry out mass-transfer operations
▪ Biochemical ‘refineries’ (liver)
• Tissue repair (wound healing)
o Wounded tissue displays a healing process that is relevant to tissue engineering
▪ A biopsied piece or graft of tissue is expected to initially display a healing-type
response after being placed in culture or engrafted.
▪ Tissue repair occurs in phases:
• Early in process (days) there is a coordination between cell proliferation,
adhesion and migration
• Remodelling of wound occurs later (weeks to years) as a result of cell
differentiation and ECM proper formation
• The healing response is faster in fetus and slower in adults
• Tissue formation (morphogenesis and development biology)
o The formation of tissue involves development biology including morphogenesis.
▪ Morphological changes are important in the formation and subsequent function
of the tissue and are fundamental to tissue formation and repair.
▪ After fertilization of an eff, several cell divisions and differentiation (is the process
where a cell changes from one cell type to another) take place. This spherical
mass reorganizes forming blastocyst containing a cavity, and starting gastrulation
which is a large-scale morphogenic process.
Morphogenesis: we have division, differentiation, migration …
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The underlying molecular-control mechanisms of morphological processes (which proceed on a
characteristic time scale) are known in detaill but they are dependent on the cell-fate processes
(division, differentiation, death, movement and adhesion) that rely on:
• direct cell-cell interactions
• cell-ECM interactions
• chemical signals
• mechanical signals
• electrical signals
Organ and tissue growth result from an integration of biophysical communication across biological
scales, both in time and space.
Tissue and organs grow to the morphology required for their function with phenomenal precision. Many
animals can even regenerate different tissues.
Cellular-fate processes
• Cell division/replication
• Cell differentiation
• Cell death
• Cell motion
• Cell adhesion
5 Common convergences between ubiquitous signalling pathways influencing tissue maturation
TF: are proteins that interact with the promoters of the gens to promote the transcription of the genes
They need to be phosphorylated by kinases. The kinases are regulated by different signals.
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Interpretation of morphogen signal/chemical signals
Morphogens (M) usually act by binding to transmembrane receptors (R) and initiating signaling
cascades, ultimately leading to the activation of morphogen effector, commonly transcription factors
(TF), which allow expression of target genes (genes X).
As a morphogen (green circle) gradient is established, naïve cells are exposed to differing
concentrations of the morphogen originating different cell types (red, white and blue). Different
concentrations of the signaling molecules lead to different molecules.
Morphogen concentration gradients are dynamics systems that can confer spatial information to cells,
which can respond to a fold change in ligand levels (i.e. rate of change), rather than absolute levels.
By analysing Smad4 activation dynamics, nodal triggers a rapid, adaptive target response dependent
on the rate of ligand concentration change. On the contrary, BMP signaling dynamics were found to
depend on absolute ligand concentration.
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It is important the type of the molecule, the concentration and the change in concentration.
Control of bone cell differentiation
Differentiation of osteoclasts
From hematopoietic stem cells (HSCs) occurs through a series og steps (osteoclastogenesis)
controlled by CSF-1 (produced by stromal cells) and RANKL, which is located on the surface of the
osteoblast (and T cells).
Differentiation of osteoblasts
From mesenchymal stem cells (MSCs) occurs through a series of steps controlled by the Wnt signalling
pathway. The osteoclasts release sclerostin (SOST), which inhibits the Wnt signalling to reduce
osteoblastogenesis. As the osteoblasts secrete bone (white arrows), they gradually become buried in
the mineralized matrix where they are transformed into star-shaped osteocytes.
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Electrical stimulation induces proliferation and differentiation in mesenchymal stem cells and
preosteoblasts.
• Electric field application leads to an increase of intracellular Ca2+ and subsequent activation of
gene expression.
• Changes in Vmem within and among cells in a network.
o Vmem is a measure of the resting
membrane potential across a
plasma membrane of a cell
created by pumps and channels
that regulate the movement of ions
across the plasma membrane.
Resting Vmem can vary within a
cell and during phases of the cell
cycle. However, the resting Vmem
is generally associated with the
proliferative state of a cell,
becoming hyperpolarized with
progressive differentiation.
o When cells are coupled in a series,
voltage differential is minimized
through ionic currents flowing
through gap junctions.
o Less differentiation cells: more positive Vmem.
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• Large-scale bioelectric patterns are instructive for shape
o Bioelectric signaling controls tissue shape and structure.
o Understanding of the bioelectric circuit controls, for example, anterior-posterior
specification in a fragment of regenerating Planaria can be used to design drug cocktails
that alter the anatomical structure thus produced, such as inducing the posterior-facing
blastema to build a secondary head in planaria.
o Depolarization of host tissues in the context of an eye transplant induces drastic over
proliferation of nerve emerging from the implanted organ in comparison to a control host.
This technique can be used to patter the ectopic nerve, inducing it to connect to specific
regions by inducing the activation o ion channels in the surrounding tissues.
Mechanical signals: Information flow and molecular origins of mechanics
• Intricacies involved with connecting molecular, cellular, and tissue scale behaviours and
mechanisms. At the molecular scale, there is molecular signaling which causes intermolecular
force production. This force production then feeds back into more molecular signaling. From the
molecular scale, there are resultant forces and cell shape changes/movements on the cellular
scale, which induce signaling into neighboring cells. The cellular scale can then feedback into
molecular scale dynamics or result in tissue scale movements or bulk mechanical property
changes. Isolating any portion of this intricate feedback loop is extremely difficult without
considering all upstream and downstream effects.
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• The different types of mechanical cues cells experience
o Fluid flow generates a shear stress parallel to the cells surface and in the direction of fluid
flow
o Compression/stretching occurs when a pushing force presses the cell inward causing it
to be compacted
o Cells are surrounded by matrix proteins. Cells respond to changes in matrix stiffness by
tuning their internal contractility
o A protrusive force is generated by the polymerization of actin at the leading edge. Actin
monomers are added at the plus end which is oriented towards the plasma membrane.
A protrusive force pushes the cell membrane forward.
o Cells forming adhesions with adjacent cells can experience a tugging face from
increased tension in the circumferential actin belt that connects the cells.
o Increased internal contractility of cells is derived from the molecular motor myosin
binding and pulling the actin filaments in opposite directions, thereby generating traction
forces.
• Integrins an cadherins modulate the mechanical landscape of the cell
o Integrin-based focal adhesions* (A) and cadherin-dependent adherens junctions (B)
relay mechanical signals through a contractile actin–myosin network (C) to actively
modulate the mechanical landscape of the cell. Focal adhesions and adherens junctions
form the linkages of the cell to the ECM and to neighboring cells, respectively. Integrins
and cadherins are linked to the intracellular actin–myosin network and are thus
intrinsically linked to each other. *Focal adhesions share some similarities with
desmosomes, although they differ in that integrins are linked to actin filaments rather
than intermediate filaments
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Intracellular mechanosensation pathways that accelerate glycolysis
• (b) Cell-cell adhesion forces are sensed through E cadherin, which forms a complex with and
activates LKB1 as well as AMPK. AMPK and the membrane cytoskeletal protein vinculin are
phosphorylated by LKB1 and Abl, respectively. Vinculin activity enhances actin remodelling
through the Rho/ROCK pathway, whereas AMPK promotes glucose uptake and ATP production
in order to maintain energy supply to adhesions.
• (d) Cell-matrix forces are sensed through integrins in focal adhesion complexes, which promote
actin fiber remodelling (also simulation of RTKs promotes actin remodelling (c)) through
Rho/ROCK and JNK/p38 signaling. Actin fibre remodelling, integrates all three signaling
pathways, and enhances transcriptional activity of YAP/TAZ as well as release of glycolytic
enzymes in the cytosol. The cytoskeleton also sequesters TRIM21 from the cytosol, preventing
proteasomal-mediated degradation of PFK. These processes culminate in acceleration of
glucose metabolism
Integrins alter cell metabolism in response to increase matrix stiffness
• (a) On soft matrices TRIM21 targets PFK1 for degradation.
o Additionally, sterol regulatory element binding proteins (SREBP) are cleaved and are then
trafficked to the nucleus. In the nucleus, SREBP signals for increased lipid synthesis
• (b) On stiff matrices, TRIM21 is sequestered by actin stress fibers, which protects PFK1 from
degradation and promotes glycolysis. The transcriptional co-activators YAP/TAX are activated by
stiff matrices and increase glutaminase (GSL1), thereby promoting glutaminolysis.
o A stiff matrix also promotes kindlin localization to the mitochondria promoting proline
synthesis
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Reciprocal regulation of mechanics and metabolism in cancer
• Under normal conditions, the compliant ECM suppresses JNK/p38signaling, and promotes
either retainment of inactive YAP/TAZ in the cytosol or its degradation by proteasomes. Cells
show reduced actin cytoskeleton assembly and TRIM21bindsto PFK promoting its degradation.
• In cancer, ECM stiffening accelerates glycolysis through JNK/p38 and YAP/TAZ mediated
metabolic reprogramming, and induces cytoskeleton remodelling, (TRIM21 becomes inactive)
resulting in the release of glycolytic enzymes in the cytosol, thereby promoting glucose
metabolism. Transcriptional programs of glucose and glutamine metabolism are promoted.
Glutamine metabolic and glycolytic end-products feed into the TCA cycle, increasing the
abundance of oncometabolites (fumarate, succinate, and 2-HG).
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Molecular connectivity from the ECM to the nucleus and nuclear membrane
mechanotransduction
• Nesprins bind to actin and to cytoskeletal cross-linkers (plectin and kinesin) and with SUN
proteins. The complex SUN-nesprins act as the physical link connecting the nucleus and the
cytoskeleton.
a) a) The nuclear envelope protein conformational changes responding to the exert force applied
on the nucleus.
b) b) Nuclear membrane stretch in response to force opens nuclear pore complexes, calcium
channels, and activates cPLA2 on the cytoplasmic side, thus increasing calcium release, import
of transcription factors (TFs), and production of arachidonic acid in the nucleoplasm.
c) c) Mechanical forces applied to the nucleus may induce chromatin opening and epigenetic
changes, that promote accessibility to TFs and regulate gene expression
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Osteogenic differentiation via mechanical and non-mechanical signals
Model of substrate mediated osteogenic differentiation wherein substrate stiffness (mechanical signal)
activates mechanotransduction pathway involving YAP/TAZ (1 A-1B) and non-mechanical signals from
substrate leads to osteogenesis via canonical BMPR signaling pathway (2).
Neurogenesis via mechanotransduction
• The nanotopography manipulates the focal adhesion signaling pathway and neurogenic
differentiation.
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