Chemosensitivity test

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Chemosensitivity , also chemosensitivity , opposite chemoresistance , individual tumor response testing ITRT , are terms for the sensitivity or insensitivity of cancer cells to a chemotherapeutic agent . Prior to individual cancer treatment, appropriate sensitivities or resistances should be recorded with in-vitro tests. These tests are currently still the subject of university research and are only used in clinics in exceptional cases.

background

Even if clinical studies determine which form of therapy is best for a patient group, the individual response of a specific patient can still only be determined after chemotherapy has started. Solid tumors in particular respond to chemotherapy in only 30% of patients. Ineffective therapies are expensive and burden the patient with toxic side effects, but have no benefit. They can also cause resistance (cross resistance) to other drugs. Chemoresistance rarely occurs for all drugs in one patient at the same time. The difficulty therefore lies in choosing the effective substances. Chemosensitivity tests could serve as a decision-making aid.

history

In the 1870s Ehrlich and Pasteur experimented with synthetic substances and substances made from microbes. They then observed the growth behavior of cultivated microbes in the presence of these manufactured substances. Ehrlich also coined the term chemotherapy and emphasized the need for substances that are selectively toxic . In 1950, Black and Spear published a study in which they examined the response of tumors in vitro using a succinate dehydrogenase- dependent color reduction test system. They found that their test system was good at predicting resistance and worse at predicting sensitivities. An improved version with a tetrazolium salt (MTT) was developed from this test system in 1983 . This MTT test was used by the National Cancer Institute in its cancer drug development and discovery program until it was superseded by the Sulforhodamine B test system.

The second important approach was also developed by Puck and Marcus in the 1950s. They used an agar-based cell culture system that preferentially supports the growth of transformed cells while non-transformed cells do not proliferate. Their findings led to the agar-based human tumor stem cell chemosensitivity test until 1977. Different terms were used for this test: clonogenic assay , human tumor stem cell (HTSC) assay , human tumor clonogenic assay or colony forming assay . The test took three weeks; Due to its complexity, it was only usable in every second case.

In the 1970s and early 1980s made it scintillation counter possible the incorporation of tritium -labeled thymidine to measure in dividing cells in cell culture systems. Radiolabeled thymidine has been used since the 1960s to utilize bacterial proliferation. The combination of agar cultures with thymidine incorporation resulted in the third test generation, which could be evaluated in 85% of the measurements over a test duration of five days. The test was developed by Kern and colleagues at the University of Los Angeles (UCLA). This procedure is still used in the CTR test.

In 2015, a chemosensitivity test based on multicellular tumor spheroids in breast cancer treated with neoadjuvant therapy was able to identify an effective drug for 95.5% of patients.

Further tests measure the vitality of the cancer cells using luminescence , fluorescence or other dyes . These include the ATP test [Kangas et al. (1984)], the DiSC test and test systems that use the dyes resazurin ( alamar blue ) or fluorescein . Methods were also worked on that could read out specific intracellular events when a cell dies ( apoptosis ).

Subdivision

The German Institute for Medical Documentation and Information (DIMDI) classifies the test systems in the 2012 updated operation and procedure code as follows:

  1. Test systems that measure cell proliferation (such as clonogenic assay, sulforhodamine B test, CTR test ...)
  2. Test systems that measure the metabolic activity of the cells (such as MTT test, ATP test ...)
  3. Test systems that measure the vitality of the cells (such as the DisC test)
  4. Other test systems (mouse models such as holofiber assay, human tumor xenograft, orthotopic and metastasis tumor models, autochthonous models)

Test steps

Four steps are the same for all systems in the implementation:

  1. Extraction and processing of living tumor tissue: Solid tumors have to be biopsied and the cells contained in them then separated. The separation takes place mechanically and enzymatically. Depending on the test system, single cell suspensions or microtumors (so-called spheroids ) are needed. Leukemia cells are isolated from blood or bone marrow, usually using density gradient centrifugation .
  2. Incubation : The tumor cells are exposed to the drugs for a few hours to weeks, usually for 4–6 days.
  3. Readout: Depending on the test system used (see above), various cellular properties are measured.
  4. Interpretation: The determined data must be evaluated and interpreted for the treatment decision. In most test systems, the drugs are classified as “potentially effective” or “ineffective” and, if necessary, at an intermediate level.

Tumors with a slow growth rate (such as breast cancer) are more difficult to test than tumors with a higher growth rate (such as ovarian cancer). The tumor tissue must not originate from necrosis and should reach an appropriate nutrient medium in good time. Contamination of the sample and the nutrient medium with microorganisms make the measurements impossible.

Problems

Since concentrations and half-lives differ significantly from drug concentrations within a tumor, tests can hardly produce concentrations comparable to those in the body. There are also huge differences between patients with the same type of tumor. In practice, all test systems are therefore based on blood plasma concentration and half-life. These are either used directly as a concentration (with different dilution levels) in the test or are used for calibration.

The issue becomes more complicated when drugs are administered as prodrugs , which are first metabolized in the body into the active drug (e.g. cyclophosphamide ). For testing, either the active form must be used, or the drug must be activated, for example, with homogenized liver extracts.

The testing of combination chemotherapies is difficult because of the synergy effects of the substances. In principle it is possible to test combination chemotherapies. However, the probably best polychemotherapy can also be composed of the most active individual drugs in the test.

Normal cell influence

Tumor tissues are heterogeneous associations consisting of fibroblasts , mesothelial cells , endothelial cells , non- transformed normal stromal cells and cancer cells . A reliable test should only determine how the drugs are affecting cancer cells. However, the neighboring cells also play an important role for the cancer cells. It is known that these cells influence the growth of cancer cells through the production of growth factors and cytokines and also that the effectiveness of drugs is changed by these three-dimensional structures. An ideal test system should try to maintain the interactions between the tumor cells and the neighboring cells, but only to read out the effects of the drugs on the tumor cells. The clonogenic assay, the CTR test, the DiSC test and the ATP test all use a test environment in which the cells cannot adhere , so that the growth of tumor cells is selectively promoted. In this environment, the neighboring cells do not divide, but retain their ability to produce cytokines and growth factors and thus support the tumor cells. The CTR test and DiSC test were developed in relation to the clonogenic assay and ATP test in such a way that the normal cell associations (spheroids) are retained.

Tumor heterogeneity

Tumors often consist of different clones that differ in terms of antigenicity , proliferation, degree of differentiation, likelihood of metastasis, etc. When tumor tissue from different locations of the same tumor is examined for chemosensitivity or resistance, differences are found in 20–30% of the examined drugs and patients. This means that the test results for solid tumors are not always representative results for all tumor cells. This may also be the reason why it is easier to predict chemoresistance than chemosensitivities.

Assumption of costs

The tests have not yet been included in the medical guidelines for the treatment of cancer patients. Statutory and private health insurances do not cover the costs unless these examinations are carried out by hospitals. Then the costs are included in the flat rate per case .

Individual evidence

  1. ↑ Andrew G Bosanquet, Sue M Richards, Rachel Wade, Monica Else, Estella Matutes, Martin JS Dyer, Saad MB Rassam, Justin Durant, Sheila M Scadding, Steve L Raper, Claire E Dearden, Daniel Catovsky: Drug cross-resistance and therapy Induced resistance in chronic lymphocytic leukaemia by an enhanced method of individualized tumor response testing . In: British Journal of Hematology . 146, No. 4, August 2009, ISSN 1365-2141 , pp. 384-395. doi : 10.1111 / j.1365-2141.2009.07741.x . PMID 19552723 .  
  2. ↑ Vincent T. DeVita, Theodore S. Lawrence, Steven A. Rosenberg, Robert A. Weinberg, Ronald A. DePinho: DeVita, Hellman, and Rosenberg's Cancer: Principles and Practice of Oncology (Cancer: Principles & Practice (DeVita)) , 8th Revised edition (REV). Edition, Lippincott Williams & Wilkins, May 1, 2008, ISBN 0-7817-7207-9 .
  3. ^ MH Tattersall, PR Harnett: Drug resistance in the clinical situation . In: Progress in Clinical and Biological Research . 223, 1986, ISSN 0361-7742 , pp. 151-159. PMID 3809197 .  
  4. ↑ Myers C, Cowan K, Sinha B, Chabner B: The phenomenon of pleiotropic drug resistance . In: Important Advances in Oncology . 1987, ISSN 0883-5896 , pp. 27-38. PMID 3331384 .  
  5. ^ WT Beck: The cell biology of multiple drug resistance . In: Biochemical Pharmacology . 36, No. 18, September 15, 1987, ISSN 0006-2952 , pp. 2879-2887. PMID 2888464 .  
  6. ↑ a b c d e JP Fruehauf, AG Bosanquet: In vitro determination of drug response: A discussion of clinical applications. . In: Principles & Practice of Oncology, PPO Updates , Volume 7. Lippincott, Philadelphia December 1, 1993, p. 16.
  7. ^ A b K. Halfter, N. Ditsch, HC Kolberg, H. Fischer, T. Hauzenberger, FE von Koch, I. Bauerfeind, G. von Minckwitz, I. Funke, A. Crispin, B. Mayer: Prospective cohort study using the breast cancer spheroid model as a predictor for response to neoadjuvant therapy - the SpheroNEO study. In: BMC Cancer . Volume 15, July 2015, p. 519, doi: 10.1186 / s12885-015-1491-7 , PMID 26169261 , PMC 4501185 (free full text).
  8. ↑ Frank Christian Kischkel, Carina Meyer, Julia Eich, Mani Nassir, Monika Mentze: Prediction of clinical response to drugs in ovarian cancer using the chemotherapy resistance test (CTR-test) . In: Journal of Ovarian Research . tape 10 , no. 1 , October 27, 2017, ISSN  1757-2215 , p. 72 , doi : 10.1186 / s13048-017-0365-9 .
  9. ^ Adrien Albert: Selective Toxicity: The Physico-Chemical Basis of Therapy , 7 Sub. Edition, Chapman & Hall, May 1985, ISBN 0-412-26010-7 .
  10. ↑ MM Black, FD Spear: Further observations on the effects of cancer chemotherapeutic agents on the in vitro dehydrogenase activity of cancer tissue. . In: Journal of the National Cancer Institute . 14, 1954, p. 1147.
  11. ^ T Mosmann: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays . In: Journal of Immunological Methods . 65, No. 1-2, December 16, 1983, ISSN 0022-1759 , pp. 55-63. PMID 6606682 .  
  12. ↑ MC Alley, DA Scudiero, A Monks, ML Hursey, MJ Czerwinski, DL Fine, BJ Abbott, JG Mayo, RH Shoemaker, MR Boyd: Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay . In: Cancer Research . 48, No. 3, February 1, 1988, ISSN 0008-5472 , pp. 589-601. PMID 3335022 .  
  13. ↑ MR Grever, SA Schepartz, BA Chabner: The National Cancer Institute: cancer drug discovery and development program . In: Seminars in Oncology . 19, No. 6, December 1992, ISSN 0093-7754 , pp. 622-638. PMID 1462164 .  
  14. ↑ Marie Suggitt, Michael C. Bibby: 50 years of preclinical anticancer drug screening: empirical to target-driven Approaches . In: Clinical Cancer Research . 11, No. 3, February 1, 2005, ISSN 1078-0432 , pp. 971-981. PMID 15709162 .  
  15. ↑ Wieland Voigt: Sulforhodamine B assay and chemosensitivity . In: Methods in Molecular Medicine . 110, 2005, ISSN 1543-1894 , pp. 39-48. doi : 10.1385 / 1-59259-869-2: 039 . PMID 15901925 .  
  16. ↑ a b c T T PUCK, PI MARCUS: Action of x-rays on mammalian cells . In: Journal of Experimental Medicine . 103, No. 5, May 1, 1956, ISSN 0022-1007 , pp. 653-666. PMID 13319584 .  
  17. ↑ AW Hamburger, SE Salmon: Primary bioassay of human tumor stem cells . In: Science . 197, No. 4302, July 29, 1977, ISSN 0036-8075 , pp. 461-463. PMID 560061 .  
  18. ↑ SE Salmon, AW Hamburger, BJ Soehnlen, BG Durie, DS Alberts, TE Moon: Quantitation of differential sensitivity of human tumor stem cells to anticancer drugs . In: The New England Journal of Medicine . 298, No. 24, June 15, 1978, ISSN 0028-4793 , pp. 1321-1327. doi : 10.1056 / NEJM197806152982401 . PMID 77475 .  
  19. ↑ AW Hamburger, SE Salmon, MB Kim, JM Trent, BJ Soehnlen, DS Alberts, HJ Schmidt: Direct cloning of human ovarian carcinoma cells in agar . In: Cancer Research . 38, No. 10, October 1978, ISSN 0008-5472 , pp. 3438-3444. PMID 210939 .  
  20. ↑ DD Von Hoff, GJ Harris, G Johnson, D Glaubiger: Initial experience with the human tumor stem cell assay system: potential and problems . In: Progress in Clinical and Biological Research . 48, 1980, ISSN 0361-7742 , pp. 113-124. PMID 7208514 .  
  21. ↑ AW Hamburger: The human tumor clonogenic assay as a model system in cell biology . In: International Journal of Cell Cloning . 5, No. 2, March 1987, ISSN 0737-1454 , pp. 89-107. doi : 10.1002 / stem.5530050202 . PMID 3553356 .  
  22. ↑ TD Brock: Bacterial growth rate in the sea: direct analysis by thymidine autoradiography . In: Science . 155, No. 3758, January 6, 1967, ISSN 0036-8075 , pp. 81-83. PMID 6015565 .  
  23. ↑ N Tanigawa, DH Kern, Y Hikasa, DL Morton: Rapid assay for evaluating the chemosensitivity of human tumors in soft agar culture . In: Cancer Research . 42, No. 6, June 1982, ISSN 0008-5472 , pp. 2159-2164. PMID 7074595 .  
  24. ↑ VK Sondak, CA Bertelsen, N Tanigawa, SU Hildebrand Zanki, DL Morton, EL grain, DH core: Clinical correlations with chemosensitivities Measured in a rapid thymidine incorporation assay . In: Cancer Research . 44, No. 4, April 1984, ISSN 0008-5472 , pp. 1725-1728. PMID 6704978 .  
  25. ↑ Efficacy test for chemotherapy. (PDF) In: Society for Biological Cancer Defense eV Accessed on June 27, 2018 .
  26. ↑ L Kangas, M Grönroos, AL Nieminen: Bioluminescence of cellular ATP: a new method for evaluating cytotoxic agents in vitro . In: Medical Biology . 62, No. 6, 1984, ISSN 0302-2137 , pp. 338-343. PMID 6543460 .  
  27. ↑ ATP test in the English language Wikipedia
  28. ↑ LM Weisenthal, JA Marsden, PL Dill, CK Macaluso: A novel dye exclusion method for testing in vitro chemosensitivity of human tumors . In: Cancer Research . 43, No. 2, February 1983, ISSN 0008-5472 , pp. 749-757. PMID 6184155 .  
  29. ↑ LM Weisenthal, PL Dill, NB Kurnick, ME Lippman: Comparison of dye exclusion assays with a clonogenic assay in the determination of drug-induced cytotoxicity . In: Cancer Research . 43, No. 1, January 1983, ISSN 0008-5472 , pp. 258-264. PMID 6571706 .  
  30. ↑ EM Larson, DJ Doughman, DS Gregerson, WF Obritsch: A new, simple, nonradioactive, nontoxic in vitro assay to monitor corneal endothelial cell viability . In: Investigative Ophthalmology & Visual Science . 38, No. 10, September 1997, ISSN 0146-0404 , pp. 1929-1933. PMID 9331256 .  
  31. ↑ GR Nakayama, MC Caton, MP Nova, Z Parandoosh: Assessment of the Alamar Blue assay for cellular growth and viability in vitro . In: Journal of Immunological Methods . 204, No. 2, May 26, 1997, ISSN 0022-1759 , pp. 205-208. PMID 9212838 .  
  32. ↑ B Rotman, C Teplitz, K Dickinson, JP Cozzolino: Individual human tumors in short-term micro-organ cultures: chemosensitivity testing by fluorescent cytoprinting . In: In Vitro Cellular & Developmental Biology: Journal of the Tissue Culture Association . 24, No. 11, November 1988, ISSN 0883-8364 , pp. 1137-1146. PMID 3192508 .  
  33. ^ R Larsson, P Nygren, M Ekberg, L Slater: Chemotherapeutic drug sensitivity testing of human leukemia cells in vitro using a semiautomated fluorometric assay . In: Leukemia . 4, No. 8, August 1990, ISSN 0887-6924 , pp. 567-571. PMID 2388483 .  
  34. ^ M Erlanson, E Daniel-Szolgay, J Carlsson: Relations between the penetration, binding and average concentration of cytostatic drugs in human tumor spheroids . In: Cancer Chemotherapy and Pharmacology . 29, No. 5, 1992, ISSN 0344-5704 , pp. 343-353. PMID 1551172 .  
  35. ↑ ES Vesell: Pharmacogenetics: multiple interactions between genes and environment as determinants of drug response . In: The American Journal of Medicine . 66, No. 2, February 1979, ISSN 0002-9343 , pp. 183-187. PMID 425962 .  
  36. ↑ RS Mehta, R Bornstein, IR Yu, RJ Parker, CE McLaren, KP Nguyen, KT Li, JP Fruehauf: Breast cancer survival and in vitro tumor response in the extreme drug resistance assay . In: Breast Cancer Research and Treatment . 66, No. 3, April 2001, ISSN 0167-6806 , pp. 225-237. PMID 11510694 .  
  37. ↑ Robert W Holloway, Rita S Mehta, Neil J Finkler, Kuo-Tung Li, Christine E McLaren, Ricardo J Parker, John P Fruehauf: Association between in vitro platinum resistance in the EDR assay and clinical outcomes for ovarian cancer patients . In: Gynecologic Oncology . 87, No. 1, October 2002, ISSN 0090-8258 , pp. 8-16. PMID 12468336 .  
  38. ^ M Cross, TM Dexter: Growth factors in development, transformation, and tumorigenesis . In: Cell . 64, No. 2, January 25, 1991, ISSN 0092-8674 , pp. 271-280. PMID 1988148 .  
  39. ↑ SU Hildebrand-Zanki, DH Kern: In vitro assays for new drug screening: comparison of a thymidine incorporation assay with the human tumor colony-forming assay . In: International Journal of Cell Cloning . 5, No. 5, September 1987, ISSN 0737-1454 , pp. 421-431. doi : 10.1002 / stem.5530050507 . PMID 3624922 .  
  40. ↑ OR Koechli, BU Sevin, JP Perras, R Angioli, A Steren, M Rodriguez, P Ganjei, HE Averette: Growth characteristics of nonmalignant cells in the ATP cell viability assay . In: Oncology . 51, No. 1, February 1994, ISSN 0030-2414 , pp. 35-41. PMID 8265101 .  
  41. ↑ DH Kern: Heterogeneity of drug resistance in human breast and ovarian cancers . In: The Cancer Journal from Scientific American . 4, No. 1, February 1998, ISSN 1081-4442 , pp. 41-45. PMID 9467045 .  
  42. ↑ JN McAlpine, SM Eisenkop, NM Spirtos: Tumor heterogeneity in ovarian cancer as demonstrated by in vitro chemoresistance assays . In: Gynecologic Oncology . 110, No. 3, September 2008, ISSN 1095-6859 , pp. 360-364. doi : 10.1016 / j.ygyno.2008.05.019 . PMID 18632143 .  
  43. ↑ MC Bird, S Forskitt, ED Gilby, AG Bosanquet: The influence of sample source and cell concentration on the in vitro chemosensitivity of haematological tumors . In: British Journal of Cancer . 53, No. 4, April 1986, ISSN 0007-0920 , pp. 539-545. PMID 3707845 .