Value of Chemoresistant Testing for Cancer Treatment Dr. Kai Schulze-Forster

Value of Chemoresistant Testingfor Cancer Treatment

Dr. Kai Schulze-Forster

.... Because humans are different!

Personalized Medicine

Introduction Introduction

The Problem 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

Biomarker Biomarker

WhichDrug?

Responder

Non-Respondersevere

side effects

Biomarker Biomarker

High need for predictive biomarkers!

There is progress especially for biologicals:

- Her2

- K-RAS mutation

- EGFR mutation

- …..

Chemoresistant Assay Chemoresistant Assay

… but for chemotherapy ??

validated biomarkers are not available

Solution: functional bioassays


Well-known method in bacterial infections:

Antibiogram

Replace bacteria by tumor cells

Oncobiogram

= Chemoresistant (Chemosensitivity) 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

96 well Microtiter plate

- 6 concentrations

- Duplicates

- Untreated control (negative control)

ABCDEFGH

1 2 3 4 5 6 7 8 9 10 11 12

D rug1 P os itive C ontro l B uffer

D rug2 N egative C on tro l



Method

Readout: ATP determination using chemoluminescence

Very sensitive: 40 cells/well

also named ATP-TCA (ATP-Tumor Chemosensitivity Assay)

LuciferaseATP + Luciferin + O2 AMP + 2P + CO2 + Light


Results: Chart

0

20

40

60

80

100

120

200 100 50 25 12,5 6,25

Test Drug Concentration %

Tu

mo

r G

row

th In

hib

itio

n %

Carboplatin

Docetaxel


Results:

Table


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


Advantages

Safe prediction of resistanceAvoidance of ineffective therapiesAvoidance of unnecessary side effects

Publications Publications

Publication search in PubMed 11/2013

Chemosensitivity Assay : 3089 entries

Chemosensitivity Testing Cancer: 534 entries

ATP-TCA: 60 entries


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. [email protected]


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).


RESULTS: RI <= 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 < 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.


Recent Results Cancer Res. 2003;161:221-30.

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. [email protected]


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.

(…)


Methods Mol Med. 2005;110:101-20.

Chemosensitivity testing using microplate adenosine triphosphate-based luminescence measurements.

Kurbacher CM, Cree IA.

Department of Gynecology and Obstertrics, University of Cologne, Cologne, Germany.


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 > 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.

Case Report Case Report

Man, 59 year old

Nasopharynx Carcinoma

April 2010 Surgery and Radiation

Mar 2011 Metastatic Prostate Ca

PSA > 500 ng/ml, Gleason 8, Start of antiandrogenic

therapy (Fugerel, LHRH)


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 PSA=296, start Second line chemotherapy

Taxotere/ Zometa/ Prednisolon


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


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

Hyperthermia 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.

Case Report HT1 Case Report HT1

Woman, 72 years old

GIST, rectum

Pretreated with Glivec




Case Report HT2 Case Report HT2

Woman, 34 years old



Summary Summary

Chemoresistant assays are useful tools to exclude non-responding chemotherapies, especially in pretreated multiresistant patients.

Zentrum für molekulare Onkologie GmbHIm Biotechnologiepark 14943 Luckenwalde

Germany (near Berlin)

Thank you

for your

attention !

Documents

Value of Chemoresistant Testing for Cancer Treatment Dr. Kai Schulze-Forster