responsibility, art & science of intentional extinction...

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Responsibility, Art & Science of Intentional Extinction, De-Extinction & Aging �17-Feb-2016 6:30-7:30 PM George Church Le Lab

LSRF

NHGRINIGMS

Azco

OppenheimerFounda6on

PersonalGenomes.org

Responsibility, Art & Science of Intentional Extinction, De-Extinction & Aging

We’ve made smallpox extinct & the opposite for 1918 flu.

What if we can preserve current biodiversity using ancient DNA?

If we can save a million lives per year by engineering wild mosquitoes, should we?

If we save lives by transplanting organs from pigs to humans and reversing human aging, will we overpopulate?

What about genetically engineering ourselves?

thebostoncalendar.com/events/george-church-phd-at-le-laboratoire-cambridge

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1974 Scientist = Enthusiast à1974 GMO à1977 Equality, Wealth-Ed-Health Cycle à1986 Animal rights à1998 hESCs à1999 Unsafe xenotransplants à 2004 Robot wars à2004 Lawyers & Synthetic DNA à2005 IRB poor data access & sharing à 2011 Gain-of-function H5N1 à2015 Patents & Germ line à 2016 Aging reversal = over population à 2016 Space radiation, 38% gravity, microbes à


Why? Precision Medicine

How? Miniaturization

What Next? Education, sharing

Engineering Genomes, Organs & Ecosystems

PersonalGenomes.org

Exponentially shrinking cost of

reading & interpreting �your genome

1990-2004 2007 2010 2013 2015

$999

$350,000

$6,000

$80 million

$3 billion

Is human genomics useful today?

Is human genomics useful today?

•  Preconception genetic counseling Tay Sachs•  In vitro fertilization diagnosis Trisomy & infertility•  Prenatal: Non-invasive testing Aneupolidy•  Newborn: screening PKU•  Child: Genetic Odyssey Beery twins•  Adolescent, Adult, Late Onset Cancer, Cardiovasc.

Genome Research à prevention/cure #1

Beery twins: “Cerebral palsy” à SPR à5HTP, DOPA gene

Genome Research à prevention/cure #2John Lauerman: Scotoma/leg pain à JAK2 à Aspirin gene

Genome Research à prevention/cure #3

Angelina Jolie: Healthy à BRCA1 à mastectomy, hysterectomy, oophorectomy

87% chance of breast cancer, 50% ovarian cancer before surgery.

5% after.

PersonalGenomes.org OpenHumans.org

Sharing data globally: Boston, Toronto, London, Ulsan, Vienna, NYC

pged.org

CongressHigh SchoolsScreenwritersMap-ed

Ting Wu



Why? Agriculture, medicine, public health

How? Delivery, efficiency, specificity.

What Next? Organs & Ecosystems

4 Mbp Radical Genome Engineering. Why?

Science 2013: Lajoie, et al. Nature 2015 Mandell et al.2. Genetic isolation

3. Metabolic Isolation

4. Multi-Virus resistance

1. Non-standard amino acids (NSAA)

Recombinase

Writing: DNA, RNA, protein

Group II intron Cas9

TALE

RecACAGE

ZnFMeganuclease

λ-bet/exo MAGE

RNA-guided CRISPR genome editing 1987: Junk DNA à 2013 Technology à Today libraries 100K+

1987Ishino,etal.(E.coli)

Human 3-Jan-13Zebrafish 29-Jan-13Yeast 4-Mar-13C. elegans 30-Jun-13Drosophila 5-Jul-13Mouse 8-Aug-13Rat 8-Aug-13Wheat 8-Aug-13Arabidopsis 20-Aug-13Rice 20-Aug-13Sorghum 2-Sep-13Tobacco 2-Sep-13Pig 6-Sep-13E. coli 29-Sep-13Silkworm 29-Oct-13Hamster 7-Jan-14Rabbit 8-Jan-14Xenopus 8-Jan-14Monkey 30-Jan-14Elephant 4-Feb-14

“Anything found to be true of E. coli must also be true of elephants.” --Jacques Monod 1954“and if not, we can make it so.” -- George Church 2014

Genetically Modified Humans

Two thousand clinical research trials. One approved: Glybera. Viral delivery. (AAV)

> $1M per dosevs $1K per genetic counseling

The most common disease

110 120

hIp://blog.stevekrause.org/2006/07/why-you-probably-outlive-average-life.html

28

Maxage

(years)

220405

19

12250

0.350.12.662100,000Adultmass(kg)

Rare, Extreme Outliers

AnAgedatabase(4212species)deMagalhaesetal.

Homo Sapiens

Cebus capucinus

Balaena mysticetus

28

Maxage

(years)

Arc%caislandica

220405

20

12250

0.350.12.662100,000Adultmass(kg)

Rare, Extreme Outliers

AnAgedatabase(4212species)deMagalhaesetal.

Heterocephalus glaber

Aging Reversal

Amy Wagers

2014 Science: Restoring systemic GDF11 levels reverses age-related dysfunction in mouse skeletal muscle. 2014 Science: Vascular and neurogenic rejuvenation of the aging mouse brain by young systemic factors. 2015 Nature Comm: Exposure to a youthful circulation rejuvenates bone repair through modulation of β-catenin

MouseSkeletalMuscle

Gomes,Sinclairetal.CellDec2013.

Nico6namide

Age-related decline in NAD+ & ATP rescued by Nicotinamide in mice

Nico6namide$1600/g0.5g/kg=$50k/day

TFAMac9va9onviaCRISPRgRNAs

Dox-inducible Cas9st1 activator integrated into murine C2C12 cells

0.5

47.5

0.4

11.3

3.71.3 1.0

0.0

10.0

20.0

30.0

40.0

50.0

60.0

TFAM

expression(Fold)

T2 T1T3T4 TSS

0.00

2.00

4.00

6.00

8.00

10.00

12.00

14.00

16.00

gRNA+Cas9-VP64

Cas9-VP64only

Blank

NAD/NADHra9o

Dhadwar et al. unpublished

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Why? Transplants, Therapy testing, personalized

How? Delivery, efficiency, specificity.

What Next? Aging reversal, BRAIN


Barthsyndrome

•  Cardiomyopathy

•  Defec6vecardiolipincomposi6on

•  TAZmuta6on

Wang,McCain,Yang,Pu,Parker,etal.NatureMedicine2014

PGP1-NT AACCATG-------------GGGACTGGGTG 0 bp PGP1-BTHH AACCATG--------------GGACTGGGTG -1 bp PGP1-NHEJ AACCATGagaagctaaccatgGGACTGGGTG +8 bp

Patient mutations à normal hiPSC à cardiomyocytes

TAZ mutation is responsible for cardiomyocyte morphology abnormality (edited PGP iPSC)

Reducedsarcomereorganiza6on--complementedbymRNA

Wang,McCain,Yang,Pu,Parker,etal.NatureMedicineMay-2014

Building human hippocampal circuits from PGP cells

4 Days 98%

Bipolar cells

Alex Ng

Hippocampal pyramidal neuronal cells

+ glial, endothelial cells

iPSC & derived neuron RNA

Yankner & Church collectiveip.com/grants/NIH:8756257

“14 IPSC lines from … late-onset AD (LOAD) and age-matched controls that have been differentiated to neurons and astrocytes. This led to the observation that IPSC-derived neural progenitors from LOAD patients differentiate to neurons prematurely, and differ in their transcriptional profiles when compared with controls.”

Tao Lu, Bruce Yankner et al Nature 2014 REST and stress resistance in ageing and Alzheimer's disease.

Rosetta BRAIN: Integration of data types Brain Research Advancing Innovative Neurotechnologies

Activity (tickertape/electrode/capillary/dust) Behavior, perception, memory (video) Connectome (synapse Ab + ID)Developmental lineage (ID)Expression (RNA, protein: ID) Serial section 4 FISSEQs: A,C,D,E

Marblestoneetal.PhysicalPrinciplesforScalableNeuralRecording.ArXiv.2013ConneconomicsbioRxiv2013.RoseIaBrainsArXiv.2014

Editing of Pig DNA May Lead to More Organs for People�Oct. 15, 2015�Carl Zimmer

Humanized pigs for transplants

Immune functions,Complement, Clotting

62 PERVs = PorcineEndogenous RetroViruses

Resistance to other viruses

ny6mes.com/2014/09/02/opinion/ways-to-reduce-the-kidney-shortage.html

Eyes

500,000deathsperyear.

4.9 million HF worldwide

20 million US sufferers

500,000 deaths per year.

Thousands

45 million blind (lens, cornea)

Science 2015Yang, Guell et al

KO62copiesofPERVs Eradica9onofthePERVtransmission

Yangetal,Science2015

32CONFIDENTIAL

PERVs

0

500

1000

1500

clone15clone20clone29clone38clone40clone41 negwater

PERVpolPERVenvPERVgag

Copynum

ber

Eradication of endogenous retroviruses



Why? Agriculture, public health, global warming

How? Gene drives

What Next? Testing, regulation

Esvelt, Smidler, Catteruccia, Church (2014) eLife

Gene drives for �Malaria,

roundworms, Lyme, Dengue, sleeping

sickness, …

Malaria & Lyme disease

life cycles

Yeast test of Gene Drives

DiCarlo, Chavez, Dietz, Esvelt, Church et al. Biorxiv Jan 2015



1974 Scientist = Enthusiast à1974 GMO à1977 Equality, Wealth-Ed-Health Cycle à1986 Animal rights à1998 hESCs à1999 Unsafe xenotransplants à 2004 Robot wars à2004 Lawyers & Synthetic DNA à2005 IRB poor data access & sharing à 2011 Gain-of-function H5N1 à2015 Patents & Germ line à 2016 Aging reversal = over population à 2016 Space radiation, 38% gravity, microbes à



1974 Scientist = Enthusiast à ELSI 36 articles. Polarized à win-win1974 GMO à 2014 Biocontainment1977 Equality, Wealth-Ed-Health Cycle à Cost reduction 1986 Animal rights à Computers & human organoids1998 hESCs à 1981 mESC, 2006 adult stem cells1999 Unsafe xenotransplants à 2015 endogenous viruses inactivated 2004 Robot wars à IGEM human practices 2004 Lawyers & Synthetic DNA à Surveillance2005 IRB poor data access & sharing à PGP & pgEd2012 Gain-of-function H5N1 à 2014 Pre-discuss Gene drives2015 Patents & Germ line à Addgene, safety+efficacy relative to standard care2016 Aging reversal = over population à Space2016 Space radiation, 38% gravity, microbes à Space genetics

SpaceGenetics.hms.harvard.edu

Church DymeckiRuvkunSinclairTabin

Wu(Director)Yankner

Space Colony Challenges

Gravity OsteoporosisNeuro-behaviorialRadiationMicrobiome …

Rare protective alleles LRP5G171V/+Extra-strongbonesMSTN-/-Leanmuscles&lowatherosclerosisSCN9A-/-Insensi6vitytopainABCC11-/-LowOdorproduc6onCCR5-/-HIVresistanceFUT2-/-NorovirusresistancePCSK9-/-LowcoronarydiseaseAPPA673T/+LowAlzheimer’sAPOEE2/E2LowAlzheimer’s(E2=R112C,R158C)GHR,GH-/-LowcancerSLC30A8-/+LowT2DiabetesIFIH1E627X/+LowT1DiabetesTERToverprod.LowAgingCDKN2Aoverprod.LowcancerTP53overprod.LowcancerGRIN2Boverprod.Highlearning&memoryPDE4Binhib.Lowanxiety,highproblemsolving(mice)

Radiation resistance improved 100,000-fold

10-fold e14-deletion50-fold recA20-fold yfjK10-fold dnaB

in E.coli, Byrne et al, eLife 2014

Extraterrestrial surgery:�Potential opportunities

Grey’s Anatomy 1:1

Contagion. Germ free Chickens & goats since 1920s.

Anesthesia. Conscious insensitivity to pain. SCN9A -/-



1974 Scientist = Enthusiast à ELSI 36 articles. Polarized à win-win1974 GMO à 2014 Biocontainment1977 Equality, Wealth-Ed-Health Cycle à Cost reduction 1986 Animal rights à Computers & human organoids1998 hESCs à 1981 mESC, 2006 adult stem cells1999 Unsafe xenotransplants à 2015 endogenous viruses inactivated 2004 Robot wars à IGEM human practices 2004 Lawyers & Synthetic DNA à Surveillance2005 IRB poor data access & sharing à PGP & pgEd2012 Gain-of-function H5N1 à 2014 Pre-discuss Gene drives2015 Patents & Germ line à Addgene, safety+efficacy relative to standard care2016 Aging reversal = over population à Space2016 Space radiation, 38% gravity, microbes à Space genetics

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