Immuno-Oncology and Breast Cancer
Scientific Realization within the PRAEGNANT Network
Peter A. Fasching
Immuno-Oncology hat vieleEinstiegsmöglichkeiten (Adams et al. 2015)
Immuntherapien
n Rekombinante Vakzinen Zytokine
n Immun – Checkpoint Inhibitorsn Zelluläre Therapien (T-Zellen / NK- Zellen)n Antikörpertherapien
n Chemotherapien Bestrahlungn Small Moleculen Hormontherapie
Radiotherapie
Chemotherapie
PRAEGANT Distribution (Basket in and basket out)
„PerFect“Pertuzumab
„Seraphina“Nab-Pclitaxel Weitere Studien
Regular BC Patients
„EMBRACA“PARP-I
„ABRAZO“PARP-I
Immun-onkologische
Studien
PRAEGANT Distribution (Basket in and basket out)
„PerFect“Pertuzumab
„Seraphina“Nab-Pclitaxel Weitere Studien
Regular BC Patients
„EMBRACA“PARP-I
„ABRAZO“PARP-I
Immun-onkologische
Studien
Immuntherapien
n Rekombinante Vakzinen Zytokine
n Immun – Checkpoint Inhibitorsn Zelluläre Therapien (T-Zellen / NK- Zellen)n Antikörpertherapien
n Chemotherapien Bestrahlungn Small Moleculen Hormontherapie
Treatments for HER2-positive Patients
Adjuvant orneoadjvuant 1st Line 2nd Line 3rd Line ≥ 4th Line
Diagnosis Metastases
TrastuzumabChemotherapy
TrastuzumabPertuzumabDocetaxel
T-DM1 No Approved Therapy
See National TherapyRecommendations
No Approved Therapy
T-DM1
Lines of Therapy for Metastatic Disease
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National Therapy Recommendations
Experimental considered
equivalent to any other treatment
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Immuntherapien
n Rekombinante Vakzinen Zytokine
n Immun – Checkpoint Inhibitorsn Zelluläre Therapien (T-Zellen / NK- Zellen)n Antikörpertherapienn Chemotherapien Bestrahlungn Small Moleculen Hormontherapie
Antibody Dependent Cellular Cytotoxicity (ADCC)
NK cells are Critical Effector Cells for Monoclonal Antibodies through ADCC
CD16’s Allele Prevalence
F/F 176 V/F 176 V/V 176
64 (42%) 76 (50%) 12 (8%)91 (50%) 71 (39%) 19 (11%)
Allelic variation analyzed in 330 normal individuals (45% African–American (top line), 55% Caucasian (bottom line)).
There is a Low Incidence of V/V in the Normal Population
Lehrnbecher et al, Blood 1999:, 94, 4220
• Patients with naturally occurring high affinity receptors for the Fc fragment havean improved outcome under Herceptin/Rituximab/Cetuximab treatment.
• Same holds true for treatment of multiple myeloma with Elotuzumab(Progression Free Survival 22.3 months vs. 9.8 months (presented at ASH 2015)
Rituximab Trastuzumab Cetuximab
Rationale for combining haNK with Antibodies
• Weng W-Ket al. J Clin Oncol 25:3712-3718, 2007
• Musolino A et al. J Clin Oncol. 2008 Apr 10;26(11)
• Zhang W J Clin Oncol 25:3712-3718, 2007
Antitumoral activity of NK cells
Downregulation of MHC IUpregulation of activating ligands
MICA, MICB
NKG2D
Ljunggren et al. 2007
Abken, Wels & Kühlcke 2014
CAR
1st Generation2nd Generation3rd Generation
T-cell receptor complex
Chimeric antigen receptors (CARs)
Genetically engineered NK-92 cellswith built-in ADCC-like activityCAR-NK-92 (taNK) The human NK cell line NK-92
as an off-the-shelf cellular therapeutic
• IL-2 dependent, continuously growing NK cell line established from a non-Hodgkin lymphoma patient in 1992
(H. Klingemann)
• Highly cytotoxic to leukemia, lymphoma and melanomadue to lack of inhibitory KIR receptors
• Safety of infusion of irradiated NK-92 demonstrated in phase I clinical studies in cancer patients with advanced
disease Tonn et al. 2001, Arai et al. 2008, Tonn et al. 2013
NK-92
Chimericantigen receptor
Target antigen
ErbB2/HER2EGFR/EGFRvIII
EpCAMGD2CD20CD19
Uherek et al. 2002Müller et al. 2008Sahm et al. 2012Esser et al. 2012
Boissel et al. 2013Schönfeld et al. 2015Romanski et al. 2016
Zhang et al. 2016Genßler et al. in press
Antitumoral activity of NK cells(Schönfeld et al. 2015)
Immuntherapien
n Rekombinante Vakzinen Zytokine
n Immun – Checkpoint Inhibitorsn Zelluläre Therapien (T-Zellen / NK- Zellen)n Antikörpertherapien
n Chemotherapien Bestrahlungn Small Moleculen Hormontherapie
Existence of an effective T-cell immuneresponse against cancer in humans
n Tumor-infiltrating T cells (TIL)are associated with betterprognosis in triple-negativebreast cancer
n Mutated tumor antigens andneo-peitopes can elicit highavidity, tumor-specific T-cellresponses
n T-cell based immunotherapyand possibly vaccinations arevery and promising newtreatment options for malignantdiseases.
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S. Adams et al. , J. Clin Oncol. 32:2959, 2014
L.B. Alexandrov et al., Nature, 2013
Prognostic relevance of tumor infiltrating lymphocytes
Mutation prevalence in tumors sorted by organs
T-Cell Cultivationfrom Tumor infiltratingLymphocytes
High quality DNA isolation from freshtumor after pathologicalassessment
DNA isolation fromblood
12
4
Sequencing
Bioinformatics and determinationof antigen-likely sequences
Synthesis of likely antigenicpeptides
6
7
8
5
3
Transfer of DNA togenotyping corefacility
B-CellCultivationfrom Blood
Transfer of peptidesto cell culture lab
9
Peptide Loading of B-Cells
10
11
Co-Cultivation of B-Cells and T-CellsMeasurement of T-CellResponse
AIMIdentification of T-cellstargeting tumor-specific
neoantigens among
tumor-infiltratinglymphocytes in triple
negative breast cancer
Working Programme
Ex vivo immune monitoring of infiltrating lymphocytes
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Antigen Fluorochrom Zellen
CK19 FITC Tumorzellen
MUC-1 PE-Cy7 Tumorzellen
CD3 V500 T-Zellen
CD4 BV421 T-Helferzellen
CD8 APC-Cy7 Zytotox.T-Zellen
FoxP3 APC Regulat.T-Zellen
CD19 PerCP B-Zellen
CD56 PE NK-Zellen
gated onlymphocytes gated onCD3+ gated onCD3+B C DA E [Ungated]MUC-1/CD3
Whole Genome Sequencing
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n 72 patients includedn 21 patients completed whole genome sequencing of tumor
and germline and RNAseqn HLA Typing prediction based on whole genome sequencing
alignment.n Results filtered based on RNA Expressionn Neoepitopes selected and ranked by
(transcripts per million) x (allele frequency of variant)
n Results: § About 40 neoepitopes predicted binders for MHC Class 1
Alleles§ About 100 neoepitopes predicted for MHC Class 2 Allleles
(to be selected to B-Cell Assays)
Neo-Epitope Aminoacid-Sequence
T-Cell response assessment in vitro
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A B C Dgated onlymphocytes gated onCD3+ gated onCD3+
mutated peptideNeo-antigen
autologous EBV-LCL
+ TILs
sort on activated T-cells
CD137
CD137
CD4
neg. HeLa-Ii HeLa-Ii/DM
CD4
IFN-γ
IFN-γ
CD
4
CD137
Flow cytometric analysis of expanded TILs (day 14)
Expansion of tumor-infiltrating T-lymphocytes
Characterization of T-cell
responses against identified tumor-specific mutations
Madan et al. 2012
Immuntherapien
n Rekombinante Vakzinen Zytokine
n Immun – Checkpoint Inhibitorsn Zelluläre Therapien (T-Zellen / NK- Zellen)n Antikörpertherapien
n Chemotherapien Bestrahlungn Small Moleculen Hormontherapie