1wt )nqtkqwu fkxgtukv[ )gpqog kp uvcdknkv[ cpf …h&0# fgec[u cv c tcvg vjcv qwijv vq jcxg ocfg...
TRANSCRIPT
![Page 1: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/1.jpg)
![Page 2: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/2.jpg)
![Page 3: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/3.jpg)
![Page 4: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/4.jpg)
O2
OH
~ 7 lesions cell / minute
![Page 5: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/5.jpg)
Consequences of Genome (in)stability
Cell survival
Cancer
Cell death
Degenerative Disease & Ageing
![Page 6: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/6.jpg)
![Page 7: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/7.jpg)
single-strand break repair nucleotide excision repair Double-strand break repair
OH
YTop1
P
5’
5’OH
YTop1
P
5’
5’P
OH
![Page 8: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/8.jpg)
End Processing
Gap Filling
Ligation
DNA Repair
Damage Detection PARP
Lig3 XRCC1
OH P PNK Polβ
5’ 3’
APTXTDP1
Lig3 XRCC1 Lig1 PCNA
Lig3 XRCC1Polβ
Lig1 PCNAPolβ Polδ/ε Fen1
Short-patch long-patch
![Page 9: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/9.jpg)
Spinocerebellar Ataxia with Axonal Neuropathy-1 (SCAN1)
cerebellar degeneration
pathology largely restricted to nervous system (no predisposition to cancer)
spinocerebellar ataxia
variable onset (~15 yrs)
SCAN1 AOA1 Normal
![Page 10: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/10.jpg)
Top2
OH
Y
5’5’
YOHP
POH
Y
Top1
P5’5’
OHY
Top1
P5’5’
P OH
El-Khamisy et al., Nature 2005
TDP1 TDP2
Ledesma & El-Khamisy et al. Nature 2009
Ahel & El-Khamisy et al., Nature 2007 El-Khamisy, EMBO Mol Med 2011
?
![Page 11: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/11.jpg)
Top2
OH
Y
5’5’
YOHP
P
TDP2
PDBs arising from TDP2 defect also cause neurodegeration
![Page 12: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/12.jpg)
PDBs arising from TDP2 defect also cause neurodegeration
TDP2
![Page 13: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/13.jpg)
Absence of TDP2 activity in Irish and Egyptian patients harbouring homozyous truncation mutations in TDP2
Y-P P
Y-P P
B UC IV-9 E B UC IV-9 E
P
Y I
* P
Y I
*
OHY
Top1
P5’5’
OHY
Top1
P5’5’
P OH
*
5’
*
3’Top2
OH
Y
5’5’
YOHP
P
TDP1TDP2
![Page 14: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/14.jpg)
Do PDBs arising from TDP2 defect also cause neurodegeration ?
> 5 0 o f t h e deregulated genes a r e a s s o c i a t e d with neurological disease in humans
![Page 15: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/15.jpg)
Gómez-Herreros, …..El-Khamisy, Nature Gent 2014
![Page 16: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/16.jpg)
Does defective PDB repair (in the nervous system) trigger hallmarks of Ageing (systemically) ?
Ageing Somatic
maintenance
Growth
GH#/#IGF'1#
Caroll, Chiang …..El-Khamisy, Hum Mol Gent 2015
![Page 17: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/17.jpg)
![Page 18: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/18.jpg)
![Page 19: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/19.jpg)
New tools for genome editing Clustered Regularly Interspaced Short Palindromic Repeats
(CRISPR)
![Page 20: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/20.jpg)
New tools for genome editing Clustered Regularly Interspaced Short Palindromic Repeats
(CRISPR)
![Page 21: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/21.jpg)
Consequences of Genome (in)stability
Cell survival
Cancer
Cell death
Ashour, Atteya, and El-Khamisy, Nature Rev Cancer 2015
![Page 22: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/22.jpg)
Damage the genome
Chemotherapy and radiotherapy
![Page 23: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/23.jpg)
Synthetic Lethality
“The exploitation of genetic defects essential for tumour cell survival by combining the defect in an affected pathway with a pharmacologically induced defect in a compensatory pathway”
![Page 24: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/24.jpg)
Pearl et al., Nature Rev Cancer. March 2015, doi:10.1038/nrc3891
DDR Cancer Signatures
![Page 25: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/25.jpg)
Pearl et al., Nature Rev Cancer. March 2015, doi:10.1038/nrc3891
DDR Cancer Signatures
Copy Number variations (CNVs)
![Page 26: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/26.jpg)
26
Success in Personalised Therapy Synthetic Lethality
HRerror-free
collapsed replication fork
PARPinhibition
PARP
SSB repair
Cell Death
Pharmaco
logic
Gen
etic
![Page 27: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/27.jpg)
BRCA2-/- sensitive to PARP inhibitors
BRCA2+/- insensitive to PARP inhibitors
In clinical use….Highly specific
No need for other DNA damaging agents PARP knockout mice are healthy
Olaparib (AZD-‐2281) has been approved by the FDA in Dec 2014
![Page 28: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/28.jpg)
The Road Ahead >>>> The Future for S.L.
Pearl et al., Nature Rev Cancer. March 2015, doi:10.1038/nrc3891
![Page 29: 1WT )NQTKQWU FKXGTUKV[ )GPQOG KP UVCDKNKV[ CPF …H&0# FGEC[U CV C TCVG VJCV QWIJV VQ JCXG OCFG VJG FGXGNQROGPV QH NKHG QP 'CTVJ KORQUUKDNG I .KPFJCN 0CVWTG O 2 OH ~ 7 lesions cell](https://reader034.vdocuments.us/reader034/viewer/2022050519/5fa341a1ed6e27281d63e94c/html5/thumbnails/29.jpg)
Thank You !
University of Sussex, UK Roger Philips, Alan Lehmann, Keith Caldecott, Louise Serpel, Penny Jeggo, Majid Hafezparast
University of Birmingham, Jo Morris
St Jude Hospital, USA Peter Mckinnon, Sachin Katyal
Newcastle University, UK Robert Lightowlers, Martin Meagher
Leiden University, Netherlands Leon Mullenders, Saskia Lagerwerf
Institute of Mol. Biology, Fleming, Greece Maria Fousteri
University of Surrey, UK Lisi Meira
Beatson Institute for Cancer Research, University of Glasgow, UK Anthony Chalmers
BARTS CRUK Peter SchmidtNHS Trust, UK Duncan Gilberts
The Netherland Cancer Institute, Amsterdam, Netherlands Conchita Vens
Drug Discovery, University of Sussex, Simon Ward Richard Angell
Institute for Biomedicine, Barcelona , Spain Travis Stracker
Yale University, USA Joann Sweasy
Past Lab Members Jessica Hudson Cornelia Meisenberg Meryem Alagoz Jean Carroll Chris Rockyard Owen Well Kay Osborn Abhishek Sharma Poorvi Patel Limei Ju
Current Lab Members Mohamed Ashour Lamia El-Shafei Reham Atteya Walaa Ramadan Waheba Elsayed Asmaa Abosoud Mohamed Kamel Menna El-Serafy
Department of Human Genetics, Nijmegen, The Netherlands. Janneke H M Schuurs-Hoeijmakers
Royal College of Surgeons, Dublin, Ireland Mark McCormack
Zewail City, Egypt Nehal Sabry Mohamed Ashour