Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors: Consensus recommendations

William T. Beck, Thomas M. Grogan, Cheryl L. Willman, Carlos Cordon-Cardo, David M. Parham, John F. Kuttesch, Michael Andreeff, Susan E. Bates, Costan W. Berard, James M. Boyett, Nathalie A. Brophy, Henk J. Broxterman, Helen S L Chan, William S. Dalton, Manfred Dietel, Antonio T. Fojo, Randy D. Gascoyne, David Head, Peter J. Houghton, Deo Kumar SrivastavaManfred Lehnert, Catherine P. Leith, Elisabeth Paietta, Zlatko P. Pavelic, Lisa Rimsza, Igor B. Roninson, Branimir I. Sikic, Peter R. Twentyman, Roger Warnke, Ronald Weinstein

Research output: Contribution to journalArticle

397 Citations (Scopus)

Abstract

Multidrug resistance (MDR), especially that associated with overexpression of MDR1 and its product, P-glycoprotein (Pgp), is thought to play a role in the outcome of therapy for some human tumors; however, a consensus conclusion has been difficult to reach, owing to the variable results published by different laboratories. Many factors appear to influence the detection of Pgp in clinical specimens, including its low and heterogeneous expression; conflicting definitions of detection end points; differences in methods of sample preparation, fixation, and analysis; use of immunological reagents with variable Pgp specificity and avidity and with different recognition epitopes; use of secondary reagents and chromogens; and differences in clinical end points. Also, mechanisms other than Pgp overexpression may contribute to clinical MDR. The combined effect of these factors is clearly important, especially among tumors with low expression of Pgp. Thus, a workshop was organized in Memphis, Tennessee, to promote the standardization of approaches to MDR1 and Pgp detection in clinical specimens. The 15 North American and European institutions that agreed to participate conducted three preworkshop trials with well-characterized MDR myeloma and carcinoma cell lines that expressed increasing amounts of Pgp. The intent was to establish standard materials and methods for a fourth trial, assays of Pgp and MDR1 in clinical specimens. The general conclusions emerging from these efforts led to a number of recommendations for future studies: (a) although detection of Pgp and MDRI is at present likely to be more reliable in leukemias and lymphomas than in solid tumors, accurate measurement of low levels of Pgp expression under most conditions remains an elusive goal; (b) tissue-specific controls, antibody controls, and standardized MDR cell lines are essential for calibrating any detection method and for subsequent analyses of clinical samples; (c) use of two or more vendor-standardized anti-Pgp antibody reagents that recognize different epitopes improves the reliability of immunological detection of Pgp; (d) sample fixation and antigen preservation must be carefully controlled; (e) multiparameter analysis is useful in clinical assays of MDR1/Pgp expression; (f) immunostaining data are best reported as staining intensity and the percentage of positive cells; and (g) arbitrary minimal cutoff points for analysis compromise the reliability of conclusions. The recommendations made by workshop participants should enhance the quality of research on the role of Pgp in clinical MDR development and provide a paradigm for investigations of other drug resistance-associated proteins.

Original languageEnglish (US)
Pages (from-to)3010-3020
Number of pages11
JournalCancer Research
Volume56
Issue number13
StatePublished - Jul 1 1996
Externally publishedYes

Fingerprint

Multiple Drug Resistance
P-Glycoprotein
Neoplasms
Epitopes
Education
Cell Line
Antibodies
Drug Resistance
Lymphoma
Leukemia

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Beck, W. T., Grogan, T. M., Willman, C. L., Cordon-Cardo, C., Parham, D. M., Kuttesch, J. F., ... Weinstein, R. (1996). Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors: Consensus recommendations. Cancer Research, 56(13), 3010-3020.

Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors : Consensus recommendations. / Beck, William T.; Grogan, Thomas M.; Willman, Cheryl L.; Cordon-Cardo, Carlos; Parham, David M.; Kuttesch, John F.; Andreeff, Michael; Bates, Susan E.; Berard, Costan W.; Boyett, James M.; Brophy, Nathalie A.; Broxterman, Henk J.; Chan, Helen S L; Dalton, William S.; Dietel, Manfred; Fojo, Antonio T.; Gascoyne, Randy D.; Head, David; Houghton, Peter J.; Srivastava, Deo Kumar; Lehnert, Manfred; Leith, Catherine P.; Paietta, Elisabeth; Pavelic, Zlatko P.; Rimsza, Lisa; Roninson, Igor B.; Sikic, Branimir I.; Twentyman, Peter R.; Warnke, Roger; Weinstein, Ronald.

In: Cancer Research, Vol. 56, No. 13, 01.07.1996, p. 3010-3020.

Research output: Contribution to journalArticle

Beck, WT, Grogan, TM, Willman, CL, Cordon-Cardo, C, Parham, DM, Kuttesch, JF, Andreeff, M, Bates, SE, Berard, CW, Boyett, JM, Brophy, NA, Broxterman, HJ, Chan, HSL, Dalton, WS, Dietel, M, Fojo, AT, Gascoyne, RD, Head, D, Houghton, PJ, Srivastava, DK, Lehnert, M, Leith, CP, Paietta, E, Pavelic, ZP, Rimsza, L, Roninson, IB, Sikic, BI, Twentyman, PR, Warnke, R & Weinstein, R 1996, 'Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors: Consensus recommendations', Cancer Research, vol. 56, no. 13, pp. 3010-3020.
Beck WT, Grogan TM, Willman CL, Cordon-Cardo C, Parham DM, Kuttesch JF et al. Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors: Consensus recommendations. Cancer Research. 1996 Jul 1;56(13):3010-3020.
Beck, William T. ; Grogan, Thomas M. ; Willman, Cheryl L. ; Cordon-Cardo, Carlos ; Parham, David M. ; Kuttesch, John F. ; Andreeff, Michael ; Bates, Susan E. ; Berard, Costan W. ; Boyett, James M. ; Brophy, Nathalie A. ; Broxterman, Henk J. ; Chan, Helen S L ; Dalton, William S. ; Dietel, Manfred ; Fojo, Antonio T. ; Gascoyne, Randy D. ; Head, David ; Houghton, Peter J. ; Srivastava, Deo Kumar ; Lehnert, Manfred ; Leith, Catherine P. ; Paietta, Elisabeth ; Pavelic, Zlatko P. ; Rimsza, Lisa ; Roninson, Igor B. ; Sikic, Branimir I. ; Twentyman, Peter R. ; Warnke, Roger ; Weinstein, Ronald. / Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors : Consensus recommendations. In: Cancer Research. 1996 ; Vol. 56, No. 13. pp. 3010-3020.
@article{d1c2fc35f2de4b4296e89651f305f925,
title = "Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors: Consensus recommendations",
abstract = "Multidrug resistance (MDR), especially that associated with overexpression of MDR1 and its product, P-glycoprotein (Pgp), is thought to play a role in the outcome of therapy for some human tumors; however, a consensus conclusion has been difficult to reach, owing to the variable results published by different laboratories. Many factors appear to influence the detection of Pgp in clinical specimens, including its low and heterogeneous expression; conflicting definitions of detection end points; differences in methods of sample preparation, fixation, and analysis; use of immunological reagents with variable Pgp specificity and avidity and with different recognition epitopes; use of secondary reagents and chromogens; and differences in clinical end points. Also, mechanisms other than Pgp overexpression may contribute to clinical MDR. The combined effect of these factors is clearly important, especially among tumors with low expression of Pgp. Thus, a workshop was organized in Memphis, Tennessee, to promote the standardization of approaches to MDR1 and Pgp detection in clinical specimens. The 15 North American and European institutions that agreed to participate conducted three preworkshop trials with well-characterized MDR myeloma and carcinoma cell lines that expressed increasing amounts of Pgp. The intent was to establish standard materials and methods for a fourth trial, assays of Pgp and MDR1 in clinical specimens. The general conclusions emerging from these efforts led to a number of recommendations for future studies: (a) although detection of Pgp and MDRI is at present likely to be more reliable in leukemias and lymphomas than in solid tumors, accurate measurement of low levels of Pgp expression under most conditions remains an elusive goal; (b) tissue-specific controls, antibody controls, and standardized MDR cell lines are essential for calibrating any detection method and for subsequent analyses of clinical samples; (c) use of two or more vendor-standardized anti-Pgp antibody reagents that recognize different epitopes improves the reliability of immunological detection of Pgp; (d) sample fixation and antigen preservation must be carefully controlled; (e) multiparameter analysis is useful in clinical assays of MDR1/Pgp expression; (f) immunostaining data are best reported as staining intensity and the percentage of positive cells; and (g) arbitrary minimal cutoff points for analysis compromise the reliability of conclusions. The recommendations made by workshop participants should enhance the quality of research on the role of Pgp in clinical MDR development and provide a paradigm for investigations of other drug resistance-associated proteins.",
author = "Beck, {William T.} and Grogan, {Thomas M.} and Willman, {Cheryl L.} and Carlos Cordon-Cardo and Parham, {David M.} and Kuttesch, {John F.} and Michael Andreeff and Bates, {Susan E.} and Berard, {Costan W.} and Boyett, {James M.} and Brophy, {Nathalie A.} and Broxterman, {Henk J.} and Chan, {Helen S L} and Dalton, {William S.} and Manfred Dietel and Fojo, {Antonio T.} and Gascoyne, {Randy D.} and David Head and Houghton, {Peter J.} and Srivastava, {Deo Kumar} and Manfred Lehnert and Leith, {Catherine P.} and Elisabeth Paietta and Pavelic, {Zlatko P.} and Lisa Rimsza and Roninson, {Igor B.} and Sikic, {Branimir I.} and Twentyman, {Peter R.} and Roger Warnke and Ronald Weinstein",
year = "1996",
month = "7",
day = "1",
language = "English (US)",
volume = "56",
pages = "3010--3020",
journal = "Journal of Cancer Research",
issn = "0099-7013",
publisher = "American Association for Cancer Research Inc.",
number = "13",

}

TY - JOUR

T1 - Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors

T2 - Consensus recommendations

AU - Beck, William T.

AU - Grogan, Thomas M.

AU - Willman, Cheryl L.

AU - Cordon-Cardo, Carlos

AU - Parham, David M.

AU - Kuttesch, John F.

AU - Andreeff, Michael

AU - Bates, Susan E.

AU - Berard, Costan W.

AU - Boyett, James M.

AU - Brophy, Nathalie A.

AU - Broxterman, Henk J.

AU - Chan, Helen S L

AU - Dalton, William S.

AU - Dietel, Manfred

AU - Fojo, Antonio T.

AU - Gascoyne, Randy D.

AU - Head, David

AU - Houghton, Peter J.

AU - Srivastava, Deo Kumar

AU - Lehnert, Manfred

AU - Leith, Catherine P.

AU - Paietta, Elisabeth

AU - Pavelic, Zlatko P.

AU - Rimsza, Lisa

AU - Roninson, Igor B.

AU - Sikic, Branimir I.

AU - Twentyman, Peter R.

AU - Warnke, Roger

AU - Weinstein, Ronald

PY - 1996/7/1

Y1 - 1996/7/1

N2 - Multidrug resistance (MDR), especially that associated with overexpression of MDR1 and its product, P-glycoprotein (Pgp), is thought to play a role in the outcome of therapy for some human tumors; however, a consensus conclusion has been difficult to reach, owing to the variable results published by different laboratories. Many factors appear to influence the detection of Pgp in clinical specimens, including its low and heterogeneous expression; conflicting definitions of detection end points; differences in methods of sample preparation, fixation, and analysis; use of immunological reagents with variable Pgp specificity and avidity and with different recognition epitopes; use of secondary reagents and chromogens; and differences in clinical end points. Also, mechanisms other than Pgp overexpression may contribute to clinical MDR. The combined effect of these factors is clearly important, especially among tumors with low expression of Pgp. Thus, a workshop was organized in Memphis, Tennessee, to promote the standardization of approaches to MDR1 and Pgp detection in clinical specimens. The 15 North American and European institutions that agreed to participate conducted three preworkshop trials with well-characterized MDR myeloma and carcinoma cell lines that expressed increasing amounts of Pgp. The intent was to establish standard materials and methods for a fourth trial, assays of Pgp and MDR1 in clinical specimens. The general conclusions emerging from these efforts led to a number of recommendations for future studies: (a) although detection of Pgp and MDRI is at present likely to be more reliable in leukemias and lymphomas than in solid tumors, accurate measurement of low levels of Pgp expression under most conditions remains an elusive goal; (b) tissue-specific controls, antibody controls, and standardized MDR cell lines are essential for calibrating any detection method and for subsequent analyses of clinical samples; (c) use of two or more vendor-standardized anti-Pgp antibody reagents that recognize different epitopes improves the reliability of immunological detection of Pgp; (d) sample fixation and antigen preservation must be carefully controlled; (e) multiparameter analysis is useful in clinical assays of MDR1/Pgp expression; (f) immunostaining data are best reported as staining intensity and the percentage of positive cells; and (g) arbitrary minimal cutoff points for analysis compromise the reliability of conclusions. The recommendations made by workshop participants should enhance the quality of research on the role of Pgp in clinical MDR development and provide a paradigm for investigations of other drug resistance-associated proteins.

AB - Multidrug resistance (MDR), especially that associated with overexpression of MDR1 and its product, P-glycoprotein (Pgp), is thought to play a role in the outcome of therapy for some human tumors; however, a consensus conclusion has been difficult to reach, owing to the variable results published by different laboratories. Many factors appear to influence the detection of Pgp in clinical specimens, including its low and heterogeneous expression; conflicting definitions of detection end points; differences in methods of sample preparation, fixation, and analysis; use of immunological reagents with variable Pgp specificity and avidity and with different recognition epitopes; use of secondary reagents and chromogens; and differences in clinical end points. Also, mechanisms other than Pgp overexpression may contribute to clinical MDR. The combined effect of these factors is clearly important, especially among tumors with low expression of Pgp. Thus, a workshop was organized in Memphis, Tennessee, to promote the standardization of approaches to MDR1 and Pgp detection in clinical specimens. The 15 North American and European institutions that agreed to participate conducted three preworkshop trials with well-characterized MDR myeloma and carcinoma cell lines that expressed increasing amounts of Pgp. The intent was to establish standard materials and methods for a fourth trial, assays of Pgp and MDR1 in clinical specimens. The general conclusions emerging from these efforts led to a number of recommendations for future studies: (a) although detection of Pgp and MDRI is at present likely to be more reliable in leukemias and lymphomas than in solid tumors, accurate measurement of low levels of Pgp expression under most conditions remains an elusive goal; (b) tissue-specific controls, antibody controls, and standardized MDR cell lines are essential for calibrating any detection method and for subsequent analyses of clinical samples; (c) use of two or more vendor-standardized anti-Pgp antibody reagents that recognize different epitopes improves the reliability of immunological detection of Pgp; (d) sample fixation and antigen preservation must be carefully controlled; (e) multiparameter analysis is useful in clinical assays of MDR1/Pgp expression; (f) immunostaining data are best reported as staining intensity and the percentage of positive cells; and (g) arbitrary minimal cutoff points for analysis compromise the reliability of conclusions. The recommendations made by workshop participants should enhance the quality of research on the role of Pgp in clinical MDR development and provide a paradigm for investigations of other drug resistance-associated proteins.

UR - http://www.scopus.com/inward/record.url?scp=8944261743&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=8944261743&partnerID=8YFLogxK

M3 - Article

C2 - 8674056

AN - SCOPUS:8944261743

VL - 56

SP - 3010

EP - 3020

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0099-7013

IS - 13

ER -