Sam Berns & Progeria: What a Rare Disease Teaches Us About a Universal Condition

Francis S. Collins, M.D., Ph.D.Director, National Institutes of Health

Thomas E. Cone Jr. History Lecture, 2015 AAP NCEOctober 24, 2015

Sam: On Living with a Rare Disease

… and Rising to Its Challenges

Clinical Picture of Hutchinson-Gilford Progeria Syndrome General:

– Short stature– Weight for height

Skin: Subcutaneous fat Hair: Alopecia Head:

– Micrognathia– Prominent scalp veins

Teeth: Delayed and misaligned Limbs:

– Coxa valga– Joints prominent and stiff

Bones: Distal resorption (clavicles, phalanges) Cardiovascular: severe atherosclerosis

A “Silent” Mutation Causes Progeria

Eriksson, M. et al., Nature (2003)

Splice donor consensus TGG A

GA G T

Normal LMNA sequence

Mutation GGC=>GGT(G608G)

G G T G G G C

G G T G G G T

3’UTR12

Normal splice: lamin A

Exon 11

progerin

Exon 10

*

*Glycine

N1 H1 N2 H2 N3 N4 HeLa no-RNA

3’UTR12

Normal: lamin A

Exon 11

progerin

Exon1--10

lamin Aprogerin

Forward primer

Reverse primer

N1-4: normal H1-2: HGPS

RT-PCR Demonstrates Abnormal Splice

Lamin A Processing

Capell B.C. & Collins F.S., Nat. Rev. Genet. (2006)

Goldman, R.D. et al., Proc. Natl. Acad. Sci. USA (2004)

Lamin AProgerinLamin C

Phenotypes in Interphase HGPS Cells

Could a Drug Block Productionof the Toxic Protein?

Capell, B.C. et al., Proc. Natl. Acad. Sci. USA (2005)

Patient Fibroblasts: In vitro 72 Hour FTI Treatment

Mouse Model of Progeria:A G608G BAC Transgenic

G608G

*P RAB25P (c1orf5)SSR2 P (FLJ12287)

164.4 kb

25.4

13.5

22.2 (24.1)

LMNA44.3

9.33.717.59.5 (11.9)

15.2

Phenotype of male double copy progeria mouse

4 mos

6 mos

8 mos

2 mos

G608G/G608G Wild type

lumen

Wild type

Progeriatransgenic

Progeriatransgenic

+ FTI

G608G BACtransgenic

showssevere lossof vascular

smooth muscle cellsin aorta at

nine months

lumen

Farnesyl-transferaseinhibitorsprevent damage

lumen

Farnesyltransferases benefit mice with progeria And kids too!

Sam on Clinical Trials

Therapeutic Strategies for Progeriaand Other Laminopathies

Small Molecule Drugs– Reduce progerin production

• Farnesyltransferase inhibitors

– Increase progerin clearance• mTOR inhibitors

RNA Therapy– Block abnormal splice

• Morpholinos

Therapeutic Strategies for Progeriaand Other Laminopathies

Small Molecule Drugs– Reduce progerin production

• Farnesyltransferase inhibitors

– Increase progerin clearance• mTOR inhibitors

RNA Therapy– Block abnormal splice

• Morpholinos

Rapamycin improves nuclear blebbing in HGPS cells

24

What about other laminopathies?

Cao et al., Science Translational Med (2011)

Next Steps for mTOR Inhibition Strategy

Trial in G608G mouse model vs. wt controls– Everolimus (mTOR inhibitor)– Lonafarnib (farnesyltransferase inhibitor)– Both drugs– Neither drug

Cross breed progeria mouse with mTOR ∆/∆ Consider human trial of lonafarnib plus everolimus

Therapeutic Strategies for Progeriaand Other Laminopathies

Small Molecule Drugs– Reduce progerin production

• Farnesyltransferase inhibitors

– Increase progerin clearance• mTOR inhibitors

RNA Therapy– Block abnormal splice

• Morpholinos

Can you deliver morpholinos to vascular smooth muscle? Sarepta eGFP tester mouse

27

Transgene only produces eGFP if morpholinoreaches tissue and blocks intronic splice donor

Morpholinoblocks splice donor

SA SD

Intravenous morpholino achieves good delivery to vascular smooth muscle cells of mouse aorta

28

vehicle only morpholino

Therapeutic Strategies for Progeriaand Other Laminopathies

Small Molecule Drugs– Reduce progerin production

• Farnesyltransferase inhibitors

– Increase progerin clearance• mTOR inhibitors

RNA Therapy– Block abnormal splice

• Morpholinos

Lots More Ideas!!

Normal aging

Is progerin expressed in normal individuals?

N1 H1 N2 H2 N3 N4 HeLa no-RNA

3’UTR12

Normal: lamin A

Exon 11

progerin

Exon1--10

lamin Aprogerin

Forward primer

Reverse primer

N1-4: normal H1-2: HGPS

RT-PCR Demonstrates Abnormal Splice

Detection of Progerin in Skin Tissues from Normal People

Major questions about role of progerin in cellular senescence

What else do we know about molecular events that lead to senescence?– Shortening of telomeres at the end of chromosomes

Does shortening of telomeres correlate with activation of progerin prodution?– YES

If telomeres are prevented from shortening (by activating telomerase), does progerin production cease?– YES

If telomeres are “uncapped” in early passage cells, does that activate progerin production?

TRF2 is a dominant negative shelterinprotein

Progerin production is upregulated in normal human cells with uncapped telomeres

Short telomeres trigger alternative splicing of dozens of genes, and activate prodution of progerin

Will a Mouse With No Progerin Be Viable?

Mouse embryo fibroblasts

Next Steps for pf/pf Mice

Detailed assessment of phenotype Assess potential for extended longevity Passage MEFs and/or skin fibroblasts repeatedly to

assess timing of senescence Compare pf/pf and wt gene expression in multiple

tissues Determine if pf/pf MEFs are protected from senescence

after telomere uncapping Determine if pf/pf background alters phenotype of

telomere KO mice

Nature is nowhere accustomed more openly to display her secret mysteries than in cases where she shows traces of her workings apart from the beaten path; nor is there any better way to advance the proper practice of medicine than to give our minds to the discovery of the usual law of nature, by the careful investigation of cases of rarer forms of disease.

~ William Harvey, Letter IX, to John Vlackveld, 24 Apr 1657

Rare Diseases: Window on Nature?

Collins LaboratoryKan CaoCecie BlairBrian CapellAmanda DuboseMike ErdosMaria ErikssonDina FaddahStephen LichtensteinNoreen PetrashUrraca TavarezMelissa TruongRenee Varga

SareptaRyszard KoleDan MourichStacy CrumleyVictoria Philbin

Progeria Res. Fndn.Leslie GordonAudrey Gordon

Dana-FarberMark Kieran

Boston Children’sMonica Kleinman

Columbia UniversityKarima Djabali

Northwestern Univ.Bob Goldman

Mass GeneralDimitri Krainc

Progeria Research Team

Sam’s Message to Researchers

Sam’s Enduring WisdomTEDxMidAtlantic 2013

NIH… Turning Discovery Into Health

directorsblog.nih.gov @NIHDirector