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Fractal circuit sensors enable rapid quantification of biomarkers for donor lung assessment for transplantation.

Sage AT, Besant JD, Mahmoudian L, Poudineh M, Bai X, Zamel R, Hsin M, Sargent EH, Cypel M, Liu M, Keshavjee S, Kelley SO - Sci Adv (2015)

Bottom Line: Using fractal circuit sensors (FraCS), three-dimensional metal structures with large surface areas, we were able to rapidly (<20 min) and reproducibly quantify small differences in the expression of interleukin-6 (IL-6), IL-10, and ATP11B mRNA in donor lung biopsies.A proof-of-concept study using 52 human donor lungs was performed to develop a model that was used to predict, with excellent sensitivity (74%) and specificity (91%), the incidence of PGD for a donor lung.This work provides an important step toward bringing rapid diagnostic mRNA profiling to clinical application in lung transplantation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 3M2, Canada.

ABSTRACT
Biomarker profiling is being rapidly incorporated in many areas of modern medical practice to improve the precision of clinical decision-making. This potential improvement, however, has not been transferred to the practice of organ assessment and transplantation because previously developed gene-profiling techniques require an extended period of time to perform, making them unsuitable in the time-sensitive organ assessment process. We sought to develop a novel class of chip-based sensors that would enable rapid analysis of tissue levels of preimplantation mRNA markers that correlate with the development of primary graft dysfunction (PGD) in recipients after transplant. Using fractal circuit sensors (FraCS), three-dimensional metal structures with large surface areas, we were able to rapidly (<20 min) and reproducibly quantify small differences in the expression of interleukin-6 (IL-6), IL-10, and ATP11B mRNA in donor lung biopsies. A proof-of-concept study using 52 human donor lungs was performed to develop a model that was used to predict, with excellent sensitivity (74%) and specificity (91%), the incidence of PGD for a donor lung. Thus, the FraCS-based approach delivers a key predictive value test that could be applied to enhance transplant patient outcomes. This work provides an important step toward bringing rapid diagnostic mRNA profiling to clinical application in lung transplantation.

No MeSH data available.


Related in: MedlinePlus

Lung transplant assessment assay.(A) The lung assessment assay workflow. A biopsy is taken from a donated lung, and the cells are homogenized and lysed. The mRNA released from the cells is analyzed using a chip-based method that delivers a gene expression profile predictive of the outcome of a transplant within 20 min of the biopsy. (B) FraCS sensor chip (left) and SEM images of sensors after electrochemical deposition (right). (C) Assay readout [PNA probe (black) and resulting differential pulse voltammetry (DPV) signal (right panel, shown in dark blue), target mRNA (light blue) hybridization, and resulting DPV signal (right panel, shown in red)].
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Figure 1: Lung transplant assessment assay.(A) The lung assessment assay workflow. A biopsy is taken from a donated lung, and the cells are homogenized and lysed. The mRNA released from the cells is analyzed using a chip-based method that delivers a gene expression profile predictive of the outcome of a transplant within 20 min of the biopsy. (B) FraCS sensor chip (left) and SEM images of sensors after electrochemical deposition (right). (C) Assay readout [PNA probe (black) and resulting differential pulse voltammetry (DPV) signal (right panel, shown in dark blue), target mRNA (light blue) hybridization, and resulting DPV signal (right panel, shown in red)].

Mentions: To bring molecular diagnostics to the bedside in LTx, we posed the critical challenge of developing an electrochemical assay that would deliver a reliable and reproducible result to the clinician in less than 30 min from sample receipt. In this limited time window, tissue must be taken from its crude state and processed into a form that can be analyzed for gene expression. Herein, we report a new class of nanoscale chip-based sensors, termed fractal circuit sensors (FraCS), which enable quantitative analysis of multiple biomarkers present at relevant concentrations in human lung tissue. The lung tissue assessment chip enabled by FraCS rapidly assesses predictive mRNA biomarkers in donor lungs from a lung tissue biopsy performed by the surgeons, relying on a simplified sample preparation step to release biomarkers into solution, thus essentially requiring no sample purification (Fig. 1A). To our knowledge, there exist no previous reports of sample-to-answer processing and analysis of lung tissue biomarkers for LTx suitability using techniques requiring fewer than 12 hours.


Fractal circuit sensors enable rapid quantification of biomarkers for donor lung assessment for transplantation.

Sage AT, Besant JD, Mahmoudian L, Poudineh M, Bai X, Zamel R, Hsin M, Sargent EH, Cypel M, Liu M, Keshavjee S, Kelley SO - Sci Adv (2015)

Lung transplant assessment assay.(A) The lung assessment assay workflow. A biopsy is taken from a donated lung, and the cells are homogenized and lysed. The mRNA released from the cells is analyzed using a chip-based method that delivers a gene expression profile predictive of the outcome of a transplant within 20 min of the biopsy. (B) FraCS sensor chip (left) and SEM images of sensors after electrochemical deposition (right). (C) Assay readout [PNA probe (black) and resulting differential pulse voltammetry (DPV) signal (right panel, shown in dark blue), target mRNA (light blue) hybridization, and resulting DPV signal (right panel, shown in red)].
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4643795&req=5

Figure 1: Lung transplant assessment assay.(A) The lung assessment assay workflow. A biopsy is taken from a donated lung, and the cells are homogenized and lysed. The mRNA released from the cells is analyzed using a chip-based method that delivers a gene expression profile predictive of the outcome of a transplant within 20 min of the biopsy. (B) FraCS sensor chip (left) and SEM images of sensors after electrochemical deposition (right). (C) Assay readout [PNA probe (black) and resulting differential pulse voltammetry (DPV) signal (right panel, shown in dark blue), target mRNA (light blue) hybridization, and resulting DPV signal (right panel, shown in red)].
Mentions: To bring molecular diagnostics to the bedside in LTx, we posed the critical challenge of developing an electrochemical assay that would deliver a reliable and reproducible result to the clinician in less than 30 min from sample receipt. In this limited time window, tissue must be taken from its crude state and processed into a form that can be analyzed for gene expression. Herein, we report a new class of nanoscale chip-based sensors, termed fractal circuit sensors (FraCS), which enable quantitative analysis of multiple biomarkers present at relevant concentrations in human lung tissue. The lung tissue assessment chip enabled by FraCS rapidly assesses predictive mRNA biomarkers in donor lungs from a lung tissue biopsy performed by the surgeons, relying on a simplified sample preparation step to release biomarkers into solution, thus essentially requiring no sample purification (Fig. 1A). To our knowledge, there exist no previous reports of sample-to-answer processing and analysis of lung tissue biomarkers for LTx suitability using techniques requiring fewer than 12 hours.

Bottom Line: Using fractal circuit sensors (FraCS), three-dimensional metal structures with large surface areas, we were able to rapidly (<20 min) and reproducibly quantify small differences in the expression of interleukin-6 (IL-6), IL-10, and ATP11B mRNA in donor lung biopsies.A proof-of-concept study using 52 human donor lungs was performed to develop a model that was used to predict, with excellent sensitivity (74%) and specificity (91%), the incidence of PGD for a donor lung.This work provides an important step toward bringing rapid diagnostic mRNA profiling to clinical application in lung transplantation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 3M2, Canada.

ABSTRACT
Biomarker profiling is being rapidly incorporated in many areas of modern medical practice to improve the precision of clinical decision-making. This potential improvement, however, has not been transferred to the practice of organ assessment and transplantation because previously developed gene-profiling techniques require an extended period of time to perform, making them unsuitable in the time-sensitive organ assessment process. We sought to develop a novel class of chip-based sensors that would enable rapid analysis of tissue levels of preimplantation mRNA markers that correlate with the development of primary graft dysfunction (PGD) in recipients after transplant. Using fractal circuit sensors (FraCS), three-dimensional metal structures with large surface areas, we were able to rapidly (<20 min) and reproducibly quantify small differences in the expression of interleukin-6 (IL-6), IL-10, and ATP11B mRNA in donor lung biopsies. A proof-of-concept study using 52 human donor lungs was performed to develop a model that was used to predict, with excellent sensitivity (74%) and specificity (91%), the incidence of PGD for a donor lung. Thus, the FraCS-based approach delivers a key predictive value test that could be applied to enhance transplant patient outcomes. This work provides an important step toward bringing rapid diagnostic mRNA profiling to clinical application in lung transplantation.

No MeSH data available.


Related in: MedlinePlus