a very basic introduction to Down syndrome, some therapeutic targets, and clinical trials sponsored by the Jerome Lejeune Foundation

1. www.lejeuneusa.org | 267-403-2910 | Contact@LejeuneUSA.org | 6397 Drexel Rd. Philadelphia, PA 19151
Pioneering innovative
therapies
toimprovethelivesofthosewithgeneticintellectualdisabilities

2. An International
Perspective on Down
Syndrome Research

3. Dr. Jerome Lejeune
1926-1994
The “Father of Modern Genetics”

4. • 1958: Discovered that Down syndrome is caused by a third copy of the 21st
chromosome. Findings published in 1959.
• 1962: Received the first Kennedy Prize
• 1964: Identified cri du chat syndrome
• 1966: Described 18q- syndrome
• 1969: Received the William Allan Memorial Award, ASHG
• 1989: Expert witness at Maryville, TN human embryo case
• 1994: Jerome Lejeune dies
• 1996: Fondation Jerome Lejeune established in Paris
• 2007: Cause for beatification and canonization is opened in Paris
• 2012: Jerome Lejeune Foundation USA established
• 2015: New Foundation office opened in Madrid and clinic in Dubai
A partial Jerome Lejeune timeline

5. Anton Van Leeuwenhoek (1632 - 1723)
• Improved microscope
• First to observe and single-cell organisms,
muscle fibers, bacteria, sperm cells, and
blood flow in capillaries
“Fathers”who changed Medicine
Jerome Lejeune (1926 - 1994)
• Discovered the genetic cause of Down
syndrome

6. Excerpted from,“Jerome Lejeune: To the least of these my brothers and sisters”
available at www.LejeuneUSA.org

7. AVERY simple biology of Down syndrome
In general
• Gene overdosing results in a metabolic disturbance that alters typical
development and results in more rapid neurological and physical
degeneration throughout the lifespan.
Fetal development
• Unregulated cell death (apoptosis), loss of neural stem cells, reduced
neurogenesis, and the failure to develop necessary communication
networks among different brain regions.
Throughout Life
• More rapid aging leading to AD symptoms and often dementia
• Other neurological conditions (catatonia, depression, oppositional
behaviors, compulsive behaviors, etc)

8. From understanding to treatment

9. InterPP (Protein - Protein Interactions)
On international open-source platform funded and created by the Jerome Lejeune
Foundation through a partnership with Hybrigenics to discover:
1. Proteins responsible for intellectual disability in Down syndrome, fragile X
syndrome and other genetic anomalies.
2. The interaction of those proteins
3. The most likely protein targets to focus on for treatment.

10. Protein Interaction Map

11. SomeTherapeuticTargets in DS Research
Restoring the metabolic imbalance caused by trisomy 21
• GABAertic system*
• Moderating gene expression
• APP*
• DYRK1A*
• CBS
• Folinic acid and thyroid*
Interesting speculative investigations that work in a lab
• XIST (Jeanne Lawrence, UMass)
• ZSCAN4 (Elixergen, LLC Baltimore)
• Sonic Hedgehog (Roger Reeves, Johns Hopkins)
Prenatal therapies
• Fluoxetine*
• Antioxidant therapies
• Choline supplementation

12. Addressing potential therapeutic targets
GABA - looking at the big picture.
Ongoing clinical trials:
•Roche, RG1662 (Basmisanil) phase 2, ages 6 - 11
•BalanceTherapeutics, BTD-001 phase 1b, ages 13-35
Rather than…

13. Neuroscience and Brain Technologies Department, Istituto Italiano di Tecnologia, Genova, Italy.
Research funded by the Jerome Lejeune Foundation
GABA - Another perspective and pathway

14. Research to address genetic expression
Single gene investigations (not comprehensive)
DYRK1A
• TesDAD (Spain)
• TRIAD (France)
APP (Amyloid precursor protein)
• The link between DS and Alzheimer’s disease?
CBS (Cystathionine-β-synthase)
• CiBleS21 Program. Jerome Lejeune Foundation
ACTHYF Project. Folinic Acid and Thyroxine (France)
• Investigations to improve psychomotor development, 6 - 18 mos.
Current/Recent clinical trials:
• EGCG, Dierssen and de la Torre (Barcelona)
• AC Immune, ACI-24 anti-amyloid vaccine
• ACTHYF, folinic acid and thyroxine. (Institut Lejeune, Paris)
• APP. Transition Therapeutics (Elan, Ireland) ELND005. Unsuccessful.

15. DYRK1A is a kinase
Kinase are responsible
for phosphorylation
Phosphorylation turns
cellular processes ON and OFF
Abnormal phosphorylation
is implicated in many diseases
PHOSPHORYLATION: The Master Switch of the Cell
DYRK1A and Kinase (VERY) basics…

16. DYRK1A Research(Dual-Specificity Tyrosine-(Y)-Phosphorylation Regulated Kinase 1A) Considered to be a strong
candidate gene for learning defects associated with Down syndrome. DYRKs have been
implicated in cell survival, proliferation and differentiation, etc.
Interactions
Phosphorylation targets

17. DYRK1A, cont…
Over expression of DYRK1A may
promote premature differentiation of
neurons and contribute to altered brain
development in those with Down
syndrome
Corticogenesis
Dierssen. Nature Reviews Neuroscience 13, 844-858 (December 2012)

18. Clinical trial sponsored by the Jerome Lejeune Foundation
Principle co-investigators: Mara Dierssen, Rafael de la Torre
Barcelona, Spain.
Epigallocatechin gallate
What is it? A natural chemical substance and antioxidant in green tea
that has been found to be an inhibitor of DYRK1A activity. In this clinical
trial EGCG was administered and accompanied by cognitive stimulation
to enhance neuroplasticity.
Result of 2014 Trial: Reported improvement in memory and executive
function. Also, evidence of normalization of plasma homocysteine
levels identifying a biomarker for testing efficacy and effect on DYRK1A.

19. Since 2014.
New clinical trial planned lowering age of subjects, improving neurostimulation,
and fine-tuning dosage.
And…

20. Leucetta microraphis
Drug development project. A partnership between the
Jerome Lejeune Foundation and ManRos Therapeutics
Roscoff in Brittany, France.
What is it? A sea sponge claimed to probably be the most selective
inhibitor of DYRK1A ever described in the scientific literature.
Goal: The molecule has been synthesized and is in development for a
phase 1 clinical trial by 2017.
TRI for Trisomy 21
AD for Alzheimer’s disease

21. Roscoff. No wonder Laurent Meijer is always smiling

22. Clinical trial designed, and carried out by the
Jerome Lejeune Institute, Paris. Funding by the
Jerome Lejeune Foundation
Principle investigator: Clotilde Mircher
What is it? A 4-arm, double-blind clinical trial to test the effect of folinic
acid and thyroxine on the psychomotor development of young children
from 6 - 18 months old. Goal, 256 subjects.
Why? There are seven genes on chromosome 21 involved in folate
metabolism. Folate deficiency can interfere with fetal and early
childhood brain development and function by restricting myelination,
dendritic arborization and synaptic connectivity.

23. The importance of folic acid and thyroid in myelination
Myelin = a fatty protein that forms the
white matter of the brain. It’s the
insulation around neurons (axons) that
assist in conduction. Think of
electrical wires and insulation.
Fluorescent images of nerve connections
in the brain of a typically-developing
individual, left, compared to a person
with Down syndrome. (Yale University)

24. We found genome-wide alterations in the expression of a large number of genes,
many of which exhibited temporal and spatial specificity and were associated with
distinct biological processes. In particular, we uncovered co-dysregulation of
genes associated with oligodendrocyte differentiation and myelination that
were validated via cross-species comparison to Ts65Dn trisomy mice. Furthermore,
we show that hypomyelination present in Ts65Dn mice is in part due to cell-
autonomous effects of trisomy on oligodendrocyte differentiation and results in
slower neocortical action potential transmission. Together, these results identify
defects in white matter development and function in DS, and they provide a
transcriptional framework for further investigating DS neuropathogenesis.
Down Syndrome Developmental Brain Transcriptome Reveals
Defective Oligodendrocyte Differentiation and Myelination.
(Neuron, Feb 25, 2016)
Jose Luis Olmos-Serrano, Hyo Jung Kang, William A. Tyler, John C. Silbereis, Feng Cheng, Ying Zhu,
Mihovil Pletikos, Lucija Jankovic-Rapan, Nathan P. Cramer, Zygmunt Galdzicki, Joseph Goodliffe, Alan
Peters, Claire Sethares, Ivana Delalle, Jeffrey A. Golden, Tarik F. Haydar, Nenad Sestan.

25. Myelination begins 14 weeks post conception and continues into early
adulthood. New research (Feb 2016) shows changes in early
development and differentiation of oligodendrocytes that produce
myelin.
WE KNOW THAT:
Folate and thyroid are critical
for optimizing myelin development
during fetal development and at
least through early childhood.
SO:
Does supplementation with folinic
acid and thyroxine rescue
myelination during infancy?
We hope the ACTHYF clinical trial
will help answer that question.

26. BUT DON’T UNDERESTIMATE THE COMPLEXITY!!!

27. Antioxidant Therapies
• Diana Bianchi,Tufts. (Apigenin)
• Maddalena Adorno, Stanford
Fluoxetine (Prozac)
• Renata Bartesaghi, Bologna
• UT Southwestern (clinical trial)
Choline Supplementation
• Barbara Strupp, Cornell
PrenatalTherapies
Diana Bianchi, MD, Clinical Geneticist
and Executive Director of the Mother
Infant Research Institute.
Tomo Tarui, MD, a pediatric neurologist
and the director of the Baby Neurology
Program
Tufts Medical Center, Boston

28. Prozac and birth defects
BMJ, 2015.
Conclusions: These data provide reassuring evidence for some SSRIs but
suggest that some birth defects occur 2-3.5 times more frequently among
the infants of women treated with paroxetine or fluoxetine early in
pregnancy. (note: This evidence of birth defects based upon a typical
population, not a DS population)

29. 6397 Drexel Rd
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Jerome Lejeune
FOUNDATION USA
Research Care Advocacy
Paris - Philadelphia - Madrid - Dubai

30. “Sonic Hedgehog Hedgehog Agonist Therapy Corrects Structural and Cognitive Deficits in a Down Syndrome Mouse
Model.” ISHITA DAS, et al. Science Translational Medicine. 04 SEP 2013 : 201RA120 http://stm.sciencemag.org/
content/5/201/201ra120
“Translating dosage compensation to trisomy 21”. Jiang J. et al. Nature. 2013 Aug 15;500 (7462):296-300. doi:
10.1038/nature12394. Epub 2013 Jul 17. http://www.ncbi.nlm.nih.gov/pubmed/23863942
“Epigallocatechin-3-gallate, a DYRK1A inhibitor, rescues cognitive deficits in Down syndrome mouse models and in
humans.” R. de la Torre, et al. Mol Nutr Food Res. 2014 Feb;58(2):278-88. doi: 10.1002/mnfr.201300325. Epub 2013
Sep 14. http://www.ncbi.nlm.nih.gov/pubmed/24039182
“Correction of Down syndrome and Edwards syndrome aneuploidies in human cell cultures”. Tomokazu Amano, et al.
DNA Res (2015) 22 (5): 331-342. doi: 10.1093/dnares/dsv016 First published online: August 31, 2015. http://
dnaresearch.oxfordjournals.org/content/22/5/331
“Effect of leucovorin (folinic acid) on the developmental quotient of children with Down's syndrome (trisomy 21) and
influence of thyroid status.” Blehaut H et al. PLoS One. 2010 Jan 11;5(1):e8394. doi: 10.1371/journal.pone.0008394.
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0008394
“Prenatal pharmacotherapy rescues brain development in a Down's syndrome mouse model.” Guidi S, et al. Brain.
2014 Feb;137(Pt 2):380-401. doi: 10.1093/brain/awt340. Epub 2013 Dec 12. http://www.ncbi.nlm.nih.gov/pubmed/
24334313
“Noninvasive prenatal testing creates an opportunity for antenatal treatment of Down syndrome.”Guedj F, Bianchi, D.
Prenat Diagn. 2013 Jun;33(6):614-8. doi: 10.1002/pd.4134. http://www.ncbi.nlm.nih.gov/pubmed/23595836. See also,
https://nihrecord.nih.gov/newsletters/2015/03_13_2015/story2.htm
“Maternal Choline Supplementation: A Potential Prenatal Treatment for Down Syndrome and Alzheimer's Disease”
Strupp BJ, et al. Curr Alzheimer Res. 2016;13(1):97-106. http://www.ncbi.nlm.nih.gov/pubmed/26391046
Short Bibliography of Signifiant Research