Novel Stool-Based Protein Biomarkers for Improved Colorectal Cancer Screening: A Case-Control Study

Linda J.W. Bosch, Meike De Wit, Thang V. Pham, Veerle M.H. Coupé, Annemieke C. Hiemstra, Sander R. Piersma, Gideon Oudgenoeg, George L. Scheffer, Sandra Mongera, Jochim Terhaar Sive Droste, Frank A. Oort, Sietze T. Van Turenhout, Ilhame Ben Larbi, Joost Louwagie, Wim Van Criekinge, Rene W.M. Van Der Hulst, Chris J.J. Mulder, Beatriz Carvalho, Remond J.A. Fijneman, Connie R. Jimenez & 1 others Gerrit A. Meijer

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Background: The fecal immunochemical test (FIT) for detecting hemoglobin is used widely for noninvasive colorectal cancer (CRC) screening, but its sensitivity leaves room for improvement.

Objective: To identify novel protein biomarkers in stool that outperform or complement hemoglobin in detecting CRC and advanced adenomas.

Design: Case-control study.

Setting: Colonoscopy-controlled referral population from several centers.

Participants: 315 stool samples from one series of 12 patients with CRC and 10 persons without colorectal neoplasia (control samples) and a second series of 81 patients with CRC, 40 with advanced adenomas, and 43 with nonadvanced adenomas, as well as 129 persons without colorectal neoplasia (control samples); 72 FIT samples from a third independent series of 14 patients with CRC, 16 with advanced adenomas, and 18 with nonadvanced adenomas, as well as 24 persons without colorectal neoplasia (control samples).

Measurements: Stool samples were analyzed by mass spectrometry. Classification and regression tree (CART) analysis and logistic regression analyses were performed to identify protein combinations that differentiated CRC or advanced adenoma from control samples. Antibody-based assays for 4 selected proteins were done on FIT samples.

Results: In total, 834 human proteins were identified, 29 of which were statistically significantly enriched in CRC versus control stool samples in both series. Combinations of 4 proteins reached sensitivities of 80% and 45% for detecting CRC and advanced adenomas, respectively, at 95% specificity, which was higher than that of hemoglobin alone (P < 0.001 and P = 0.003, respectively). Selected proteins could be measured in small sample volumes used in FIT-based screening programs and discriminated between CRC and control samples (P < 0.001).

Limitation: Lack of availability of antibodies prohibited validation of the top protein combinations in FIT samples.

Conclusion: Mass spectrometry of stool samples identified novel candidate protein biomarkers for CRC screening. Several protein combinations outperformed hemoglobin in discriminating CRC or advanced adenoma from control samples. Proof of concept that such proteins can be detected with antibody-based assays in small sample volumes indicates the potential of these biomarkers to be applied in population screening.

Primary Funding Source: Center for Translational Molecular Medicine, International Translational Cancer Research Dream Team, Stand Up to Cancer (American Association for Cancer Research and the Dutch Cancer Society), Dutch Digestive Foundation, and VU University Medical Center.

Original languageEnglish
Pages (from-to)855-866
Number of pages12
JournalAnnals of Internal Medicine
Volume167
Issue number12
Early online date21 Nov 2017
DOIs
Publication statusPublished - 19 Dec 2017

Cite this

Bosch, Linda J.W. ; De Wit, Meike ; Pham, Thang V. ; Coupé, Veerle M.H. ; Hiemstra, Annemieke C. ; Piersma, Sander R. ; Oudgenoeg, Gideon ; Scheffer, George L. ; Mongera, Sandra ; Droste, Jochim Terhaar Sive ; Oort, Frank A. ; Van Turenhout, Sietze T. ; Larbi, Ilhame Ben ; Louwagie, Joost ; Van Criekinge, Wim ; Van Der Hulst, Rene W.M. ; Mulder, Chris J.J. ; Carvalho, Beatriz ; Fijneman, Remond J.A. ; Jimenez, Connie R. ; Meijer, Gerrit A. / Novel Stool-Based Protein Biomarkers for Improved Colorectal Cancer Screening : A Case-Control Study. In: Annals of Internal Medicine. 2017 ; Vol. 167, No. 12. pp. 855-866.
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abstract = "Background: The fecal immunochemical test (FIT) for detecting hemoglobin is used widely for noninvasive colorectal cancer (CRC) screening, but its sensitivity leaves room for improvement.Objective: To identify novel protein biomarkers in stool that outperform or complement hemoglobin in detecting CRC and advanced adenomas.Design: Case-control study.Setting: Colonoscopy-controlled referral population from several centers.Participants: 315 stool samples from one series of 12 patients with CRC and 10 persons without colorectal neoplasia (control samples) and a second series of 81 patients with CRC, 40 with advanced adenomas, and 43 with nonadvanced adenomas, as well as 129 persons without colorectal neoplasia (control samples); 72 FIT samples from a third independent series of 14 patients with CRC, 16 with advanced adenomas, and 18 with nonadvanced adenomas, as well as 24 persons without colorectal neoplasia (control samples).Measurements: Stool samples were analyzed by mass spectrometry. Classification and regression tree (CART) analysis and logistic regression analyses were performed to identify protein combinations that differentiated CRC or advanced adenoma from control samples. Antibody-based assays for 4 selected proteins were done on FIT samples.Results: In total, 834 human proteins were identified, 29 of which were statistically significantly enriched in CRC versus control stool samples in both series. Combinations of 4 proteins reached sensitivities of 80{\%} and 45{\%} for detecting CRC and advanced adenomas, respectively, at 95{\%} specificity, which was higher than that of hemoglobin alone (P < 0.001 and P = 0.003, respectively). Selected proteins could be measured in small sample volumes used in FIT-based screening programs and discriminated between CRC and control samples (P < 0.001).Limitation: Lack of availability of antibodies prohibited validation of the top protein combinations in FIT samples.Conclusion: Mass spectrometry of stool samples identified novel candidate protein biomarkers for CRC screening. Several protein combinations outperformed hemoglobin in discriminating CRC or advanced adenoma from control samples. Proof of concept that such proteins can be detected with antibody-based assays in small sample volumes indicates the potential of these biomarkers to be applied in population screening.Primary Funding Source: Center for Translational Molecular Medicine, International Translational Cancer Research Dream Team, Stand Up to Cancer (American Association for Cancer Research and the Dutch Cancer Society), Dutch Digestive Foundation, and VU University Medical Center.",
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author = "Bosch, {Linda J.W.} and {De Wit}, Meike and Pham, {Thang V.} and Coup{\'e}, {Veerle M.H.} and Hiemstra, {Annemieke C.} and Piersma, {Sander R.} and Gideon Oudgenoeg and Scheffer, {George L.} and Sandra Mongera and Droste, {Jochim Terhaar Sive} and Oort, {Frank A.} and {Van Turenhout}, {Sietze T.} and Larbi, {Ilhame Ben} and Joost Louwagie and {Van Criekinge}, Wim and {Van Der Hulst}, {Rene W.M.} and Mulder, {Chris J.J.} and Beatriz Carvalho and Fijneman, {Remond J.A.} and Jimenez, {Connie R.} and Meijer, {Gerrit A.}",
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Bosch, LJW, De Wit, M, Pham, TV, Coupé, VMH, Hiemstra, AC, Piersma, SR, Oudgenoeg, G, Scheffer, GL, Mongera, S, Droste, JTS, Oort, FA, Van Turenhout, ST, Larbi, IB, Louwagie, J, Van Criekinge, W, Van Der Hulst, RWM, Mulder, CJJ, Carvalho, B, Fijneman, RJA, Jimenez, CR & Meijer, GA 2017, 'Novel Stool-Based Protein Biomarkers for Improved Colorectal Cancer Screening: A Case-Control Study' Annals of Internal Medicine, vol. 167, no. 12, pp. 855-866. https://doi.org/10.7326/M17-1068

Novel Stool-Based Protein Biomarkers for Improved Colorectal Cancer Screening : A Case-Control Study. / Bosch, Linda J.W.; De Wit, Meike; Pham, Thang V.; Coupé, Veerle M.H.; Hiemstra, Annemieke C.; Piersma, Sander R.; Oudgenoeg, Gideon; Scheffer, George L.; Mongera, Sandra; Droste, Jochim Terhaar Sive; Oort, Frank A.; Van Turenhout, Sietze T.; Larbi, Ilhame Ben; Louwagie, Joost; Van Criekinge, Wim; Van Der Hulst, Rene W.M.; Mulder, Chris J.J.; Carvalho, Beatriz; Fijneman, Remond J.A.; Jimenez, Connie R.; Meijer, Gerrit A.

In: Annals of Internal Medicine, Vol. 167, No. 12, 19.12.2017, p. 855-866.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Novel Stool-Based Protein Biomarkers for Improved Colorectal Cancer Screening

T2 - A Case-Control Study

AU - Bosch, Linda J.W.

AU - De Wit, Meike

AU - Pham, Thang V.

AU - Coupé, Veerle M.H.

AU - Hiemstra, Annemieke C.

AU - Piersma, Sander R.

AU - Oudgenoeg, Gideon

AU - Scheffer, George L.

AU - Mongera, Sandra

AU - Droste, Jochim Terhaar Sive

AU - Oort, Frank A.

AU - Van Turenhout, Sietze T.

AU - Larbi, Ilhame Ben

AU - Louwagie, Joost

AU - Van Criekinge, Wim

AU - Van Der Hulst, Rene W.M.

AU - Mulder, Chris J.J.

AU - Carvalho, Beatriz

AU - Fijneman, Remond J.A.

AU - Jimenez, Connie R.

AU - Meijer, Gerrit A.

PY - 2017/12/19

Y1 - 2017/12/19

N2 - Background: The fecal immunochemical test (FIT) for detecting hemoglobin is used widely for noninvasive colorectal cancer (CRC) screening, but its sensitivity leaves room for improvement.Objective: To identify novel protein biomarkers in stool that outperform or complement hemoglobin in detecting CRC and advanced adenomas.Design: Case-control study.Setting: Colonoscopy-controlled referral population from several centers.Participants: 315 stool samples from one series of 12 patients with CRC and 10 persons without colorectal neoplasia (control samples) and a second series of 81 patients with CRC, 40 with advanced adenomas, and 43 with nonadvanced adenomas, as well as 129 persons without colorectal neoplasia (control samples); 72 FIT samples from a third independent series of 14 patients with CRC, 16 with advanced adenomas, and 18 with nonadvanced adenomas, as well as 24 persons without colorectal neoplasia (control samples).Measurements: Stool samples were analyzed by mass spectrometry. Classification and regression tree (CART) analysis and logistic regression analyses were performed to identify protein combinations that differentiated CRC or advanced adenoma from control samples. Antibody-based assays for 4 selected proteins were done on FIT samples.Results: In total, 834 human proteins were identified, 29 of which were statistically significantly enriched in CRC versus control stool samples in both series. Combinations of 4 proteins reached sensitivities of 80% and 45% for detecting CRC and advanced adenomas, respectively, at 95% specificity, which was higher than that of hemoglobin alone (P < 0.001 and P = 0.003, respectively). Selected proteins could be measured in small sample volumes used in FIT-based screening programs and discriminated between CRC and control samples (P < 0.001).Limitation: Lack of availability of antibodies prohibited validation of the top protein combinations in FIT samples.Conclusion: Mass spectrometry of stool samples identified novel candidate protein biomarkers for CRC screening. Several protein combinations outperformed hemoglobin in discriminating CRC or advanced adenoma from control samples. Proof of concept that such proteins can be detected with antibody-based assays in small sample volumes indicates the potential of these biomarkers to be applied in population screening.Primary Funding Source: Center for Translational Molecular Medicine, International Translational Cancer Research Dream Team, Stand Up to Cancer (American Association for Cancer Research and the Dutch Cancer Society), Dutch Digestive Foundation, and VU University Medical Center.

AB - Background: The fecal immunochemical test (FIT) for detecting hemoglobin is used widely for noninvasive colorectal cancer (CRC) screening, but its sensitivity leaves room for improvement.Objective: To identify novel protein biomarkers in stool that outperform or complement hemoglobin in detecting CRC and advanced adenomas.Design: Case-control study.Setting: Colonoscopy-controlled referral population from several centers.Participants: 315 stool samples from one series of 12 patients with CRC and 10 persons without colorectal neoplasia (control samples) and a second series of 81 patients with CRC, 40 with advanced adenomas, and 43 with nonadvanced adenomas, as well as 129 persons without colorectal neoplasia (control samples); 72 FIT samples from a third independent series of 14 patients with CRC, 16 with advanced adenomas, and 18 with nonadvanced adenomas, as well as 24 persons without colorectal neoplasia (control samples).Measurements: Stool samples were analyzed by mass spectrometry. Classification and regression tree (CART) analysis and logistic regression analyses were performed to identify protein combinations that differentiated CRC or advanced adenoma from control samples. Antibody-based assays for 4 selected proteins were done on FIT samples.Results: In total, 834 human proteins were identified, 29 of which were statistically significantly enriched in CRC versus control stool samples in both series. Combinations of 4 proteins reached sensitivities of 80% and 45% for detecting CRC and advanced adenomas, respectively, at 95% specificity, which was higher than that of hemoglobin alone (P < 0.001 and P = 0.003, respectively). Selected proteins could be measured in small sample volumes used in FIT-based screening programs and discriminated between CRC and control samples (P < 0.001).Limitation: Lack of availability of antibodies prohibited validation of the top protein combinations in FIT samples.Conclusion: Mass spectrometry of stool samples identified novel candidate protein biomarkers for CRC screening. Several protein combinations outperformed hemoglobin in discriminating CRC or advanced adenoma from control samples. Proof of concept that such proteins can be detected with antibody-based assays in small sample volumes indicates the potential of these biomarkers to be applied in population screening.Primary Funding Source: Center for Translational Molecular Medicine, International Translational Cancer Research Dream Team, Stand Up to Cancer (American Association for Cancer Research and the Dutch Cancer Society), Dutch Digestive Foundation, and VU University Medical Center.

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DO - 10.7326/M17-1068

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EP - 866

JO - Annals of Internal Medicine

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SN - 0003-4819

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