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. 2021 Jun 30;13(13):3303.
doi: 10.3390/cancers13133303.

A Novel Automated Immunoassay Platform to Evaluate the Association of Adiponectin and Leptin Levels with Breast Cancer Risk

Debora Macis  1 , Valentina Aristarco  1 , Harriet Johansson  1 , Aliana Guerrieri-Gonzaga  1 , Sara Raimondi  2 , Matteo Lazzeroni  1 , Ivana Sestak  3 , Jack Cuzick  3 , Andrea DeCensi  3   4 , Bernardo Bonanni  1 , Sara Gandini  2
Affiliations

A Novel Automated Immunoassay Platform to Evaluate the Association of Adiponectin and Leptin Levels with Breast Cancer Risk

Debora Macis et al. Cancers (Basel). .

Abstract

Adiponectin and leptin are adipokines secreted by the adipose tissue that are associated with several chronic diseases including cancer. We aimed to compare the immunoassay platform ELLA with an enzyme-linked immunosorbent assay (ELISA) kit and to assess whether the results of the association analyses with breast cancer risk were dependent on the assay used. We measured adiponectin and leptin with ELLA and ELISA on baseline serum samples of 116 Italian postmenopausal women enrolled in two international breast cancer prevention trials. Results were compared with Deming, Passing-Bablok regression and Bland-Altman plots. Disease-free survival was analyzed with the Cox model. There was a good correlation between the methods for adiponectin and leptin (r > 0.96). We found an increased breast cancer risk for very low adiponectin levels (HR for ELLA = 3.75; 95% CI: 1.37;10.25, p = 0.01), whereas no significant association was found for leptin levels. The disease-free survival curves were almost identical for values obtained with the two methods, for both biomarkers. The ELLA platform showed a good concordance with ELISA for adiponectin and leptin measurements. Our results support the association of very low adiponectin levels with postmenopausal breast cancer risk, irrespective of the method used. The ELLA platform is a time-saving system with high reproducibility, therefore we recommend its use for biomarker assessment.

Keywords: ELISA; ELLA; adiponectin; breast cancer risk; immunoassay platform; leptin.

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Conflict of interest statement

J.C. received funding for IBIS-II study from Cancer Research UK and AstraZeneca. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results. The other authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Correlation between ELLA (3-step dilution protocol) and ELISA for adiponectin measurement (n = 46). (A) Passing–Bablok and Pearson’s r coefficient, (B) Deming regression, (C) Bland–Altman plot (differences vs. average), (D) Bland–Altman plot (ratio vs. average), (E) Bland–Altman plot (differences vs. average) for adiponectin < 6.75 µg/mL (<first quartile Q1), and (F) for adiponectin ≥ 6.75 µg/mL (≥first quartile, Q2-3-4). Dashed lines represent 95% LoA.
Figure 2
Figure 2
Correlation between ELLA (2-step dilution protocol) and ELISA for adiponectin measurement (n = 95). (A) Passing–Bablok and Pearson’s r coefficient, (B) Deming regression, (C) Bland–Altman plot (differences vs. average), (D) Bland–Altman plot (ratio vs. average), (E) Bland–Altman plot (differences vs. average) for adiponectin < 6.75 µg/mL (<first quartile Q1), and (F) for adiponectin ≥ 6.75 µg/mL (≥first quartile, Q2-3-4). Dashed lines represent 95% LoA.
Figure 3
Figure 3
Correlation between ELLA and ELISA for leptin measurement (n = 104). (A) Passing–Bablok and Pearson’s r coefficient, (B) Deming regression, (C) Bland–Altman plot (differences vs. average), (D) Bland–Altman plot (ratio vs. average), (E) Bland–Altman plot (differences vs. average) for leptin < 33 ng/mL (<third quartile, Q1-2-3), and (F) for leptin ≥ 33 ng/mL (≥third quartile, Q4). Dashed lines represent 95% LoA.
Figure 4
Figure 4
Disease-free survival curves according to lowest quartile versus higher for adiponectin (n = 116) measured with ELLA (A) or ELISA (B) and highest quartile versus lower for leptin (n = 104) measured with ELLA (C) or ELISA (D). Disease-free survival was calculated including subjects with resected DCIS and high risk healthy subjects.
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