5. Norddt. Zytostatikaworkshop HH

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Value of Chemoresistant Testingfor Cancer
Treatment
Dr. Kai Schulze-Forster
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Introduction
Personalized Medicine
.... Because humans are different!
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The Problem
The response to chemotherapy (solid tumors) is
about 30 ! Principles and Practice of Oncology
(Cancer Principles Practice (DeVita, 8th
Revised edition (REV).Lippincott Williams
Wilkins, 1. Mai 2008
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Biomarker
WhichDrug?
Responder
severeside effects
Non-Responder
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Biomarker
High need for predictive biomarkers!
There is progress especially for biologicals-
Her2- K-RAS mutation- EGFR mutation - ..
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Chemoresistant Assay
but for chemotherapy ?? validated biomarkers
are not available Solution functional bioassays
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Chemoresistant Assay
Well-known method in bacterial infections Antibio
gram Replace bacteria by tumor cells Oncobiogram
Chemoresistant (Chemosensitivity) Assay
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Chemoresistant Assay
Method 1. Collect tumor cells (from primary
tumor, metastases or ascites during surgery,
biopsy or punction) 2. Prepare single cell
suspension 3. Perform proliferation assay with
drugs 1-3 days 4. Determine number of living
cells 5. Calculate growth inhibition
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Chemoresistant Assay

• 96 well Microtiter plate
• 6 concentrations
• Duplicates
• Untreated control (negative control)

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Chemoresistant Assay
Method Readout ATP determination using
chemoluminescence Very sensitive 40
cells/well also named ATP-TCA (ATP-Tumor
Chemosensitivity Assay)
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Chemoresistant Assay
Results Chart
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Chemoresistant Assay
Results Table
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Chemoresistant Assay
Advantages 1. In vitro-testing does not affect
the patient 2. several drugs and combinations can
be tested in parallel 3. hyperthermia can be
included
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Chemoresistant Assay
Advantages

• Safe prediction of resistance
• Avoidance of ineffective therapies
• Avoidance of unnecessary side effects

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Publications
Publication search in PubMed 11/2013

• Chemosensitivity Assay 3089 entries
• Chemosensitivity Testing Cancer 534 entries
• ATP-TCA 60 entries

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Publications
Cancer Chemotherapy and Pharmacology May 2012,
Volume 69, Issue 5, pp 1307-1314 Predicting
platinum resistance in primary advanced ovarian
cancer patients with an in vitro resistance
index Thea Eline Hetland, Janne Kærn, Martina
Skrede, Berit Sandstad, Claes Tropé, Ben
Davidson,Vivi Ann Flørenes Departement of
Gynecologic Oncology, Norwegian Radium Hospital,
Oslo University Hospital, 0424 Oslo, Norway.
theaeline_at_gmail.com
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Publications
PURPOSE We aimed to identify primary platinum
resistance in epithelial ovarian cancer (OC)
patients with FIGO stage III-IV disease by an in
vitro drug-response assay and to correlate the
findings with clinical response. () METHODS We
combined the ATP-based tumor-chemosensitivity and
the extreme drug resistance assays for testing of
85 biopsies from 58 patients. Tumors were
classified as sensitive or resistant by a
resistance index (RI). () Results were analyzed
for association with clinical platinum
resistance, progression-free survival (PFS), and
overall survival (OS).
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Publications
RESULTS RI lt 250 predicted primary platinum
resistance, without misclassification of
sensitive patients. The test sensitivity for
primary tumors was 15/15, specificity 3/10,
negative predictive value 3/3, and positive
predictive value 15/22. Patients with in vitro
platinum-resistant samples had shorter PFS
compared with patients with sensitive samples
(3.4 vs. 10.0 months, p 0.02). Comparing
patient-matched primary and metastatic samples,
there was about 1/3 mismatch in resistance. RI
for platinum was lower in primary tumors exposed
to neoadjuvant chemotherapy than in chemo-naïve
tumors (p lt 0.01). CONCLUSIONS This in vitro
assay predicted primary platinum resistance,
without misclassification of sensitive OC
patients, and the results were significantly
associated with PFS. We suggest that samples from
primary tumor and metastatic samples have
different responses to chemotherapy and that
exposure to chemotherapy might induce in vitro
platinum resistance.
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Publications
Recent Results Cancer Res. 2003161221-30. ATP
chemosensitivity testing in ovarian and breast
cancer early clinical trials. Kurbacher CM,
Grecu OM, Stier U, Gilster TJ, Janát MM, Untch M,
Konecny G, Bruckner HW, Cree IA. Division of
Clinical and Experimental Gynecologic Oncology,
Department of Gynecology and Obstetrics,
University of Cologne Medical Center, Kerpener
Strasse 34, 50931 Köln, Germany.
Christian.Kurbacher_at_medizin.uni-koeln.de
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Publications
ABSTRACT () Currently, the ATP-based tumor
chemosensitivity assay (ATP-TCA) can be regarded
as the most sophisticated assay to investigate
both solid samples and effusions derived from
patients with various organ tumors. ()Clinical
trials that have been set up in heavily
pretreated patients with recurrent ovarian or
breast cancer have convincingly confirmed the
high activity of these combinations previously
demonstrated in preclinical investigations using
the ATP-TCA. In a recent phase II trial performed
in 59 patients with relapsed ovarian carcinoma,
ATP-TCA-directed therapy was able to triple the
response rate and to double the survival time,
compared with published empirical chemotherapy
regimes. ()
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Publications
Methods Mol Med. 2005110101-20. Chemosensitivit
y testing using microplate adenosine
triphosphate-based luminescence
measurements. Kurbacher CM, Cree IA. Department
of Gynecology and Obstertrics, University of
Cologne, Cologne, Germany.
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Publications
ABSTRACT () Among these, the ATP-TCA has gained
particular merits for ex vivo chemosensitivity
testing of native nonhematological tumors
including cancers of the breast, ovary,
gastrointestinal tract, cervix and corpus uteri,
and lung malignant melanomas gliomas sarcomas
and mesotheliomas. For this indication, the
ATP-TCA can now be considered the best documented
and validated technology. ()In ovarian and
breast carcinomas, the predictive accuracy is gt
90, with a positive predictive value of 85-90
and a negative predictive value near 100,
respectively. In primary ovarian cancers, the
ATP-TCA has been found to accurately predict both
clinical response and survival.
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Case Report
Man, 59 year oldNasopharynx Carcinoma April 2010
Surgery and Radiation Mar 2011 Metastatic
Prostate CaPSA gt 500 ng/ml, Gleason 8, Start of
antiandrogenic therapy (Fugerel, LHRH)
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Case Report
April 2011 Estracyt, Zometa, PSA decreased to
21, good general condition May-July 2011 Zometa
and hormons, PSA 9 Aug 2011 Zometa, well-being,
PSA 22 Oct 2011 PSA296, start Second line
chemotherapy Taxotere/ Zometa/ Prednisolon
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Case Report
Dec 2011 Therapy well tolerated, EPO Jan 2012
Relapse Nasopharynx Ca Feb 2012 Chemoresistant
assay Cisplatin / 5-FU / Taxans found as
effective drugs Mar 2012 Start chemotherapy
(Cisplatin/5-FU/Taxotere) Aug 2012 After 6 cycles
good general condition
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Case Report
Sep 2012 Progression of Nasopharynx Ca Oct 2012
Patient dead Summary using the chemoresistant
assay a well tolerated chemotherapy could be
found for a multiple treated patient. About 6
month of life prolongation
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Hyperthermia
To simulate a hyperthermia we included a 1 hour
treatment at 42 C at incubation start. In
parallel the same drugs were tested without
hyperthermia.
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Case Report HT1
Woman, 72 years old GIST, rectum Pretreated with
Glivec
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Hyperthermia
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Hyperthermia
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Hyperthermia
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Case Report HT2
Woman, 34 years old
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Hyperthermia
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Hyperthermia
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Summary
Chemoresistant assays are useful tools to exclude
non-responding chemotherapies, especially in
pretreated multiresistant patients.
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Zentrum für molekulare Onkologie GmbH Im
Biotechnologiepark ? 14943 Luckenwalde Germany
(near Berlin)
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Thank youfor yourattention !