Stem Cells and Diabetes

Professor Anthony Gavalas

Stem Cells in Pancreas Regeneration
Pancreas : structure and function
 Diabetes : a complex chronic degenerative disease

Type I : Insulin producing beta cells are lost due to T-cell mediated
autoimmune destruction - polygenic

Type II : Relative insulin deficiency usually due to reduced insulin sensitivity

of peripheral tissues - polygenic

MODY: Autosomal dominant mutations that disrupt β cell function

monogenic
(Gck, but mostly developmental regulators such as Pdx1, Hnf4a, Hnf1a)

The difficulty of matching insulin replacement to the rapidly fluctuating

demands causes the complications of diabetes

Transplantations of whole pancreas or islets provide a cure but immunosupression

and tissue availability limit these options.

Can stem cells provide an alternative therapy ?

Paths to restoration of β cell mass
Embryonic Stem cells vs Somatic Stem cells
EMBRYONIC STEM CELLS
Self renewing, pluripotent cells derived from blastocyst stage embryos

Unique features
They can grow in culture for long periods of time
They retain normal karyotype
They can generate every cell type in the body
Differentiate in vitro recapitulating the in vivo differentiation process

BUT ...
Develop antigenicity upon differentiation
Low efficiency, mixed lineages of in vitro differentiation
ADULT STEM CELLS
Self renewing, pluripotential cells that are tissue specific
They have the ADVANTAGE that they are

Patient’s own cells, therefore there is no immune rejection

BUT ...
Low amounts in vivo
Limited regenerative capacity
Slow growth in vitro
INDUCED PLURIPOTENT STEM CELLS (iPS)
Generated from somatic cells through reprogramming
to an embryonic - like state
Pancreas development
Future endoderm cells ingress form the anterior primitive streak …

… in a temporal order prefiguring their anteroposterior position

Nodal signaling determines the endoderm (high) to mesoderm (medium) choice

Pancreas inductive signals
Gut Regionalization : Signals, Genes and Territories
Lineage specification in pancreas development
 S1p

S1pr2

Notch HIGH Notch LOW Gai Ga12

Duct Ngn3 YAP

Acinar
lineage lineage

Endocrine
lineage
From hPS cells to beta cells in seven complex steps
Encapsulation technologies to protect hPS cell derived islets

! reduce / eliminate fibrotic response

! promote vascularisation
! provide transient oxygenation
! open with immunosuppression ?

! engineering challenges
! open device
! immunosuppression
! low C-peptide (30-40 pM in 3, < 10 in all others
> 1000 pM after islet transplantation
200 pM some glycemic contol
5-30 pM reduced no of hyperglycemic events
 Nov 17, 2021

… a critical reading
https://www.healthnewsreview.org/2021/11/cure-for-type-1-diabetes-weve-been-down-this-road-before-with-ny-times/
Fetal and mature pancreas – function and cell populations
Possible sources for replenishing beta cell mass
What was shown …
Activation of Ngn3 expression correlated with increased b cell numbers
Ngn3+ cells originated from hormone- cells close to the ducts
Ngn3+ cells can be purified from the injured pancreas …
… and differentiate into functional islet cells in vitro

… and what was not shown …

Direct lineage relation of Ngn3+ cells to newly formed islet cells
The cells of origin of the Ngn3+ cells
Low efficiency of conversion

Viral vectors inappropriate for therapeutic approaches

What was shown …
All types of pancreatic cells are derived from CA II expressing cells postnatally

Following PDL, CAII expressing cells gave rise to beta cells and acinar cells

… and what was not shown …

Promoter leakiness / expression during development not adequately addressed

Are all ductal cells capable to transdifferentiate or a subset ?

 What was shown …
Adult duct cells can be converted into beta cells
This conversion was mediated by Ngn3 stabilisation

… and what was not shown …

Is there a subpopulation competent to do that ?
Fbw7 is a tumour suppressor … its inactivation may lead to serious side effects
 What was shown …

Aldh+ cells gives rise to endocrine and acinar cells in vitro and in embryonic explants

Aldh+ cells expand after pancreatic injury

… and what was not shown …

Aldh activity can be attributed to 18 different genes … which one is at play here ?

No in vivo evidence
Aldh1b1 is expressed in the emerging pancreatic buds and all progenitors
 Aldh1b1+ centroacinar cells convert to all three lineages

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Aldh1b1CreERT2 ROSA26 STOP tdTomato Aldh1b1CreERT2/ROSA26 STOP tdTomato