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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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Responsibility, Art & Science of Intentional Extinction, De-Extinction & Aging
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
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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?
• 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 #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
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Engineering Genomes, Organs & Ecosystems
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
Engineering Genomes, Organs & Ecosystems
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
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
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Engineering Genomes, Organs & Ecosystems
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, …
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Responsibility, Art & Science of Intentional Extinction, De-Extinction & Aging
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
----- Reading -------------- Writing --------- ArithmELSI -----
Responsibility, Art & Science of Intentional Extinction, De-Extinction & Aging
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
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 -/-
----- Reading -------------- Writing --------- ArithmELSI -----
Responsibility, Art & Science of Intentional Extinction, De-Extinction & Aging
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