New models for studies on metastatic cascades and therapy efficacy in

melanoma brain metastases

Herrera Ríos, Dayana “Vascular Oncology & Metastasis”

Skin Cancer Unit of the Dermatology Department University Hospital Essen, Germany

Folie 2 Titel11.10.17

MelanomaIncidence

• in2008>200.000newcasesworldwide;88.500(49%)ineurope

• calculationfor2025->350.000newcases/yearworldwide

1

2

3

Folie 3 Titel11.10.17

Brainmetastasesinmelanomapatients

• numberofBMdirectsOSinmelanomapatients

• mediansurvivalofmelanomapatientswithBM~3.8month

• BMincidence10-40%duringlifetimeBUT~75%onpatientsautopsy

Thereisaclearneedforimprovedsystemictherapies!

Folie 4 Titel11.10.17

Immune checkpoints

Cytokine Growth Factor Rev. 2017, 36:33-38

Ipilimumab

NivolumabInhibitors

Folie 5 Titel11.10.17

MT/ret:Aspontaneoustumormodelformalignantmelanoma

Nature Reviews Cancer 7, 35-45 (January 2007)

Ø fusion of human ret proto-oncogene to mouse metallothionein (MT)-I promotor

Ø ret overexpression promotes melanin production in the root sheets of hair follicles

Ø activation of MAPK signaling pathway

Ø Induction of melanoma in C57Bl/6

ret

MT/ret

MT- I

• Kalkavan Hetal.,Naturecommun 2017• KleinJCetal.,Naturecommun 2017• Carpinteiro etal.,EMBOMolMed.2015• Baldetal.,Nature2014• Helfrichetal.,PCMR2014• Hansenetal.,JExpMed 2013• Helfrichetal.,JExpMed 2010

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Primary melanoma Lymph node

Tyrosinase/PI

MT/ret:Aspontaneousmelanomamodelforbrainmetastasis

Spleen mets

fl-VCT

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TUMORPROGRESSION

QuantificationoftumorloadbyComputerTomography

Metastaticseedingcorrelateswithtumorload

Folie 8 Titel11.10.17

• Establishmentofcelllinesfrombrainmets• Analyzingbraincolonization&organhomingafterretransplantation

Generationof“matched”celllines

Brainmetastasis

Skinmelanoma

Metastaticlymphnode

BM-ME5

CM

LN

ü RNASeq profileü Functional analysisü Firstinvivoassays are running

Well-characterizedsyngeneicanimalmodelsforBM

Folie 9 Titel11.10.17

ME-CM(skinmelanoma)

melaninremovalbyCRISP/Cas9technology

Fluorochromlabeling

Winkler/Berghoff

CMcloneinducesbrainmacrometastasis

7differentclones

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Metastatic potential

Cancer Lett. 2017, 10;391:125-140.

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BrainmetcelllineRNAexpressionofEMT

MelanomaB16

Skinmelanoma

Melanomalymphnodes

Melanomabrainmet

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• CM(generatedfromprimarymelanoma)

• BM-ME5(generatedfrombrainmet)

Invivoimagingofbrainpenetrationofmelanomacelllines

Ctrl: PBS

Ctrl: PBS

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Summary

• Generated new cell lines with potential for macro metastases for syngeneic transplantation into immunosuppressed mice

• Cell lines, generated from spontaneous brain mets for studies on tumor cell homing

• Development of treatment strategies that increase cancer immune checkpoint therapy

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Group „Vascular Oncology & Metastasis“

Verena SchmittKarolin WieberStefanie LöffekIsa WestedtMara EilebrechtDirk SchadendorfIris Helfrich

Department of Neurology

Josephine Herz

F. WinklerA. Berghoff

Funding:

Acknowledgements

Dr. Konstanze Wegener Stiftung

G. ReifenbergerJ. Felsberg

G. SchackertM. Kirsch

M. HölzelD. Hinze

M. Kato