Neuroblastoma is a tumour arising from anomalies in the development of the sympathic nervous system and still accounts for 13% of all cancer-related death in children due to resistant, relapsing and metastatic diseases. There is an urgent need for the development of new treatment against high-risk relapsed neuroblastoma. Overview: Here we will discuss the ICR Paediatric Mouse Hospital approach which integrates more advanced mouse modelling, such as the use of genetically-engineered mouse (GEM) models and patient-derived xenografts to accelerate the discovery and evaluation of novel therapeutic strategies and help shape the clinical trial pipeline priorities for children with high-risk relapsing/refractory neuroblastoma. We will also highlight the pivotal role of MRI within the Mouse Hospital which includes: Enhancing and accelerating preclinical trials Quantitatively inform on tumour phenotype and tumour response to treatment to: Develop in vivo models that emulate the clinical treatment resistant phenotype using chemotherapy-dose escalation protocol Characterize tumour spatial heterogeneity and evolution over treatment and guide the pathological and molecular characterization of the resistant phenotype Finally we will also discuss how the compact, cryogen-free and user-friendly Aspect Imaging M-Series has transformed our way of working within the mouse hospital by providing a shared and easily accessible resource for tumour screening (with minimal onboarding) .

1. Accelerating the Delivery of
New Treatments for Children
with Neuroblastoma
with the MRI-Guided
Hospital
Dr. Evon Poon
Senior Scientist
The Institute of Cancer Research,
London
Dr. Yann Jamin
Application Specialist,
Scintica

2. Accelerating the Delivery of New Treatments for
Children with Neuroblastoma with the MRI-Guided
Paediatric Mouse Hospital
• Part 1 : ICR Paediatric Mouse Hospital and preclinical trials of
new therapeutics strategies for the children with
neuroblastoma
• Part 2 : The practical aspects and challenges of implementing
and running MRI-embedded preclinical trials

3. Accelerating the Delivery of New Treatments for Children with
Neuroblastoma with the MRI-Guided Paediatric Mouse Hospital
Evon Poon
Chesler Lab,
Institute of Cancer Research, UK

5. Precision Medicine for High-Risk Neuroblastoma
The Mouse Hospital and Co-Clinical Trial Approach
Multiplex imaging
Immuno-proteomics
Regional and Cell Classification
Spatial transcriptomic
Genomic
Non invasive Imaging
• The application of the mouse hospital and co-clinical trial concept represents a clear paradigm shift
in NB translational research.
• This approach integrates more advanced mouse modelling to accelerate the discovery and
evaluation of novel therapeutic strategies
• Integrate imaging and all these information to better understand the resistance and heterogeneity.

6. Precision Medicine for High-Risk Neuroblastoma
The Mouse Hospital and Co-Clinical Trial Approach
Multiplex imaging
Immuno-proteomics
Regional and Cell Classification
Spatial transcriptomic
Genomic
Non invasive Imaging
Novel models
(iPSC)
Immunology
models
Transgenic mice
modelling genetic
drivers
PDX
models
Chemoresistant
models

7. Neuroblastoma – A neural crest tumour
• Sympathetic nerve and adrenal
gland tumours
• Most common tumour after brain
• Causes 15% of childhood deaths
• The older you are the worse it is
(>18 months)
• Hereditary and sporadic forms
MYCN amp
diploid MYCN

8. Clinical characteristics of neuroblastoma
Fischer, Science 2018
• Transgenic mice modelling
genetic drivers such as
MYCN, p53, RAS, ALK, etc

9. GEMM for Pre-clinical Trials in Neuroblastoma
Th-MYCN GEM Model
Th-MYCN: Increase in MYCN dosage augments
tumorigenesis. (Weiss et al, 1997)
Human MYCN cDNA
Tyrosine-OH promotor
Reviewed in Chesler, SCB, 2011, CCR 2013
Tissue specific promotor (Tyr-OH) restricted gene overexpression in neural crest
• workhorse model, MYCN-dependent, recapitulates
all major features and 2o genomics.
• Trials: 14-21 day single-agent trials on 1o tumour
with surrogate imaging (MRI).

10. Th-MYCN GEM Model
Undifferentiated neuroblastoma
50 mm
Th-MYCN
mouse
A robust platform to evaluate novel therapy and imaging biomarkers
• The most established model of NB, which emulates the major genomic
and histopathological hallmarks of high-risk MYCN-amplified disease.

11. Neuroblastoma GEM models (subclass driver)
x
Th-MYCN/Th-ALKF1174L (Chesler, 2012) Th-MYCN/Trp53KI (Chesler, 2016)
x
Th promoter ALKF1174L
Th promoter MYCN
Th-MYCN
Th promoter MYCN
Th-MYCN
Th-ALKF1174L
x

12. Targeting MYCN degradation in neuroblastoma
Poon, Chesler, Buechel and Eilers Labs
• MYCN-amplified neuroblastoma cells express elevated
levels of Aurora-A.
• Aurora-A associates with MYCN (N-Myc) and prevents
Fbxw7-mediated proteasomal degradation of MYCN.
• Aurora A inhibitors dissociate AURKA from MYCN
• Aurora A inhibitors downregulate MYCN

13. Targeting MYCN degradation in neuroblastoma
• Aurora A inhibitors decrease colony formation and proliferation of
MYCN-amplified NB cells

14. Multimodality imaging driven trials
LHS
Kidney
RHS
Kidney RHS
Kidney
LHS
Kidney
IAUGC60 0.13 ± 0.01mM Gd min
RHS
Kidney
LHS
Kidney
Anatomic
Volumetric
(T1 vol)
-/+10%
Functional
(DCE-MRI)
RHS
Kidney
Day 0
randomise
(N=4/arm)
Precision Medicine for High-Risk Neuroblastoma
The Mouse Hospital and Co-Clinical Trial Approach

15. Multimodality imaging driven trials
RHS
Kidney
Volumetric
(T1 vol)
-/+10%
RHS
Kidney
Day 0
randomise
(N=4/arm)
Precision Medicine for High-Risk Neuroblastoma
The Mouse Hospital and Co-Clinical Trial Approach
Day 7

16. Targeting MYCN degradation in neuroblastoma
MLN8054 MLN8237
• Aurora A inhibitors regress tumours and extend survival of Th-MYCN mice.

17. Targeting MYCN degradation in neuroblastoma
• Tumours relapse while still on treatment

18. Targeting transcription-replication conflict in
neuroblastoma
• Aurora A interacts with MYCN at S-phase to prevent
transcription-replication stress
• Induction of transcription-replication conflicts opens a wide
therapeutic window
• Aurora A inhibitors activate ATR
• Combination of Aurora-A and ATR inhibitors cures a subset
of mice

19. Transgenic mice
modelling genetic
drivers
PDX
models
Novel models
(neural crest cell)
Chemoresistant
models
Immunology
models
Th-MYCN GEM Model
A robust platform to evaluate novel therapy and imaging biomarkers

20. Audience Poll

21. The practical aspects and challenges of implementing
and running MRI-embedded preclinical trials
Yann Jamin
Children with Cancer Research Fellow
Centre for Cancer Imaging
Division of Radiotherapy and Imaging
Institute of Cancer Research, UK

22. The Centre for Cancer Imaging
@ The Institute of Cancer Research London
7T MRI
SPECT/PET/CT
mCT
High-frequency US
Photo-acoustics
Optical CT
“The Centre for Cancer Imaging is a
leading edge preclinical research
facility that brings together multi-
disciplinary research teams with an
ethos of collaboration and
innovation. Its core purpose is to
develop and implement state-of-the-
art non-invasive imaging
technologies in order to support the
discovery and development of
personalized cancer therapies and
ultimately deliver improved
outcomes for cancer patients.”

23. 23
 Adult glioma (o.t)
 Pediatric glioma (o.t)
 Medulloblastoma (GEMM)
 Craniopharyngioma (GEMM)
 Multiple Myeloma (o.t.)
 Prostate bone metastasis (o.t)
Breast cancer (o.t.)
Pancreatic cancer (o.t., GEMM)
Neuroblastoma (GEMM)
Colon cancer metastasis
(o.t., PD organoids) …
Prostate cancer ( GEMMM)
Wilm’s tumour (o.t.)
Supporting the research community at ICR
Fat suppressed T2-weighted anatomical MRI

24. The Neuroblastoma
Mouse Hospital
Undifferentiated neuroblastoma
50 mm
Th-MYCN mouse
A robust platform to evaluate novel
therapy and imaging biomarkers

25. The Neuroblastoma
Mouse Hospital
A robust platform to evaluate novel
therapy and imaging biomarkers
Diagnosis
10 Jan
After 2 cycles of
chemotherapy
3 Mar
Temozolomide
9 months old

26. The Neuroblastoma
Mouse Hospital
Preclinical Trial Design
Screening
 Palpation
 T2w-MRI
DOB Genotyping
Palpation
~ Size 5
Hemizygous
Th-MYCN+/-
Homozygous
Th-MYCN+/+
Penetrance 25%
Doubling time ~7days
Penetrance 100%
Doubling time ~3days
Th-MYCN+/+

27. The Neuroblastoma
Mouse Hospital
Preclinical Trial Design
Screening
 Palpation
 T2w-MRI
DOB Genotyping
Palpation
~ Size 5
Survival trial
D0 D3
MRI MRI
D1: Cdk2/9 inh
PK/PD
Path
Intervention
trial
Cdk2/9 inh 1 and inh 2
Cdk2/9 inh 1
Cdk2/9 inh 2

28. Imaging data sets – Tumour volume segmentation

29. Imaging data sets – Tumour volume segmentation
• Dicom format: 2-3Mb/data sets
• Many software available for segmentation

30. The Neuroblastoma
Mouse Hospital
Preclinical Trial Design
Screening
 Palpation
 T2w-MRI
DOB Genotyping
Palpation
~ Size 5
Survival trial
D0 D3
MRI MRI
D1: Cdk2/9 inh
PK/PD
Path
Intervention
trial
Cdk2/9 inh 1 and inh 2
Cdk2/9 inh 1
Cdk2/9 inh 2

31. The Neuroblastoma
Mouse Hospital
Preclinical Trial Design
Total mice for single drug efficacy studies:
• Palpation-guided: 32
• MRI-guided: 16
• Increase accuracy: Compared to physical palpation
• Reduced bias: Compared to physical palpation
• Reduction in animal used: Enhanced statistical power
• Enhanced data curation

32. The Neuroblastoma
Mouse Hospital
Preclinical Trial Design
Screening
 Palpation
 T2w-MRI
DOB Genotyping
Palpation
~ Size 5
Survival trial
D0 D3
MRI MRI
D1: Cdk2/9 inh
PK/PD
Path
Intervention
trial
Cdk2/9 inh 1 and inh 2
Cdk2/9 inh 1
Cdk2/9 inh 2

33. Chemoresistant neuroblastoma provides new insights into
chemorefractory disease and metastasis
Yogev et al., 2019
• Th-MYCN mice develop
cyclophosphamide resistance following
individualized multicycle treatment.
• Resistant Th-MYCN tumours acquire genomic changes
reflective of human neuroblastoma.

34. Establishing a Screening Facility
Behind the Clean Barrier
M-SeriesTM Compact MRI
Main surgical procedure room
Drug trials animal
holding room
M-SeriesTM Compact MRI
o Permanent magnet :
o No cryogen or water needed for cooling
o Normal electrical sockets
o Fully-shielded
o Silent scanning for T2-weighted images
o Low running and maintenance cost

35. Establishing a Screening Facility
Behind the Clean Barrier
M-SeriesTM Compact MRI
Main surgical procedure room
Drug trials animal
holding room
M-SeriesTM Compact MRI
o Permanent magnet :
o No cryogen or water needed for cooling
o Normal electrical sockets
o Fully-shielded
o Silent scanning for T2-weighted images
o Low running and maintenance cost

36. Establishing a Screening Facility
Behind the Clean Barrier
M-SeriesTM Compact MRI
• Accessible to anyone
o Following 20 minutes onboarding
o No worries about quenching, or metals !
o Streamline processes
o Integrated physiological maintenance and
monitoring
o Simple acquisition software

37. Establishing a Screening Facility
Behind the Clean Barrier
M-SeriesTM Compact MRI
1 Tesla

38. Establishing a Screening Facility
Behind the Clean Barrier
M-SeriesTM Compact MRI
7 Tesla
1 Tesla

39. Establishing a Screening Facility
Behind the Clean Barrier
M-SeriesTM Compact MRI
Tumour
k
k
Acquisition time 6 min 3 min 1 min 30 s 50 s
7 Tesla
1 Tesla

40. Establishing a Screening Facility
Behind the Clean Barrier
M-SeriesTM Compact MRI
O.t. brain tumour
Fat-suppressed T2 weighted: FSE (TE/TR=10-60/4000ms, Inversion time = 100ms)
Th-MYCN GEMM
2min scan 8min scan
TRAMP GEMM
8min scan
3min scan
KPC GEMM
O.t. Prostate bone marrow metastasis
12min scan
RH-41 xenograft
4min scan
10min scan

41. WWW.SCINTICA.COM
Footer 41
41
WWW.SCINTICA.COM
M-Series
Compact, Cryogen-free, Self-shielded 1T
MRI system

42. Conclusions
• MRI is a pivotal component of the Pediatric Mouse Hospital at ICR
• MRI is helping accelerate the delivery of preclinical trials of novel
treatment against neuroblastoma.
• Aspect Imaging M-SeriesTM a cost-effective, user-friendly compact
system that improves access to MRI to the wider cancer research
community

43. Acknowledgements
Chesler lab:
Louis Chesler
Karen Barker
Barbara Martins da Costa
Kevin Greenslade
Sara Heuss
Kate Liodaki
Andrea Lampis
Claire Lynn
Lizzie Tucker
Quinty Vellema
Giuseppe Barone
Paul Workman
MRI (ICR):
Yann Jamin
Simon Robinson
Alumni:
Orli Yogev
Libby Calton
Federica Lorenzi
Collaborators:
Sally George (ICR) Karen Liu (KCL)
Franki Spelemen (Ghent) John Anderson (UCL)
Walter Koach (Dublin) Karin Straathof (UCL)
Anna Philpott (Cambridge)
Anestis Tsakiridis (Sheffield)
Martin Eilers, Gabriele Buechel (Wurzburg)
Charles Lin, Tong Liang (BCM)

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