Before clinical trials for cognition in Down syndrome:

Challenges & approaches in model systems

Katheleen Gardiner, PhD NDSC, Denver, July 18, 2013

Challenges for model systems

DS trisomy of chromosome 21
160 protein genes plus 300-400 RNA and other genes

3 chr21 vs. 2 chr21

increased levels of proteins produced from 160 genes proteins interact with each other

Goals for model systems

- not just to understand what each chr21 protein does

Understand how the many proteins of chr21 work together to produce cognitive deficits Learn how to manipulate the effects of trisomy 21 to lessen/prevent cognitive deficits
pharmacology

Model systems
Whole organisms
Mouse Zebrafish Fruit fly Brewers yeast

Human cell lines

White blood cell lines Reprogrammed, neuronal cell lines

Mouse as a model system

Mmu16 Chr21q

Chr21 genes are dispersed within

pieces of 3 chromosomes in mouse - creaEng a complete trisomy chr21 mouse is not pracEcal currently

Mmu17 160 protein genes

100 chr21 protein genes

Mmu10
20 chr21 protein genes 40 chr21 protein genes

The most popular DS mouse model Ts65Dn

Strengths Several features relevant to DS
learning/memory deficits reduced sizes of some brain regions reduced cell numbers in some brain regions abnormal cell size/shape

Several drug treatments correct abnormalities Weaknesses Partial trisomy mouse chr16 region only
Three copies of only 88 chr21 protein genes

Three copies of 50 non-chr21 genes

Successful drug treatments in Ts65Dn

Adult mice ~15 different drugs/small molecules, with very different properties
Antioxidants Anti-inflammatory agents Prozac Melatonin Enhancement of neurotransmitter functions Inhibition of neurotransmitter functions Green tea extract

Clinical trials so far no signicant improvement

Why might human clinical trials fail?

Based on success with the Ts65Dn mouse Limitations of Ts65Dn:
Does not have 3 copies of 72 of 160 chr21 protein genes
absent trisomy of genes known to influence learning/ memory, brain development, brain structure/function

Plus 3 copies of 50 non-chr21 genes Limited genetic diversity

An additional, human, model system

- iPSCs induced pluripotent stem cell lines
Human skin or blood cells
Re-program to neuronal (brain) cells

How are DS cells different from control cells? How do DS cells differ from each other? How do drugs affect DS cells?
Do all drugs produce the same results? Does a drug affect all DS cells the same way? Are there correlations with cognitive strengths & weaknesses of donors?

Comparing model systems

Mouse All chr21 genes? Genetic diversity Drug effects on neuronal structure Drug effects on learning & memory Time to test 10-20 drugs Time to test drug combinations Expense to test 10-20 drugs No No Yes Yes Years Years High Human iPSCs Yes Yes Yes No Months Months Moderate

Conclusions
Mouse is the only model system for testing of drug effects on learning & memory Human cell lines allow testing of drug effects on complete trisomy of chr21, with genetic diversity, and for correlation of cell features with human cognitive variation Human clinical trials are expensive in time, effort and $$$