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      MetaNorm: incorporating meta-analytic priors into normalization of NanoString nCounter data

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      Bioinformatics
      Oxford University Press

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          Abstract

          Motivation

          Non-informative or diffuse prior distributions are widely employed in Bayesian data analysis to maintain objectivity. However, when meaningful prior information exists and can be identified, using an informative prior distribution to accurately reflect current knowledge may lead to superior outcomes and great efficiency.

          Results

          We propose MetaNorm, a Bayesian algorithm for normalizing NanoString nCounter gene expression data. MetaNorm is based on RCRnorm, a powerful method designed under an integrated series of hierarchical models that allow various sources of error to be explained by different types of probes in the nCounter system. However, a lack of accurate prior information, weak computational efficiency, and instability of estimates that sometimes occur weakens the approach despite its impressive performance. MetaNorm employs priors carefully constructed from a rigorous meta-analysis to leverage information from large public data. Combined with additional algorithmic enhancements, MetaNorm improves RCRnorm by yielding more stable estimation of normalized values, better convergence diagnostics and superior computational efficiency.

          Availability and implementation

          R Code for replicating the meta-analysis and the normalization function can be found at github.com/jbarth216/MetaNorm.

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          Most cited references17

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          Direct multiplexed measurement of gene expression with color-coded probe pairs.

          We describe a technology, the NanoString nCounter gene expression system, which captures and counts individual mRNA transcripts. Advantages over existing platforms include direct measurement of mRNA expression levels without enzymatic reactions or bias, sensitivity coupled with high multiplex capability, and digital readout. Experiments performed on 509 human genes yielded a replicate correlation coefficient of 0.999, a detection limit between 0.1 fM and 0.5 fM, and a linear dynamic range of over 500-fold. Comparison of the NanoString nCounter gene expression system with microarrays and TaqMan PCR demonstrated that the nCounter system is more sensitive than microarrays and similar in sensitivity to real-time PCR. Finally, a comparison of transcript levels for 21 genes across seven samples measured by the nCounter system and SYBR Green real-time PCR demonstrated similar patterns of gene expression at all transcript levels.
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            Normalization of RNA-seq data using factor analysis of control genes or samples.

            Normalization of RNA-sequencing (RNA-seq) data has proven essential to ensure accurate inference of expression levels. Here, we show that usual normalization approaches mostly account for sequencing depth and fail to correct for library preparation and other more complex unwanted technical effects. We evaluate the performance of the External RNA Control Consortium (ERCC) spike-in controls and investigate the possibility of using them directly for normalization. We show that the spike-ins are not reliable enough to be used in standard global-scaling or regression-based normalization procedures. We propose a normalization strategy, called remove unwanted variation (RUV), that adjusts for nuisance technical effects by performing factor analysis on suitable sets of control genes (e.g., ERCC spike-ins) or samples (e.g., replicate libraries). Our approach leads to more accurate estimates of expression fold-changes and tests of differential expression compared to state-of-the-art normalization methods. In particular, RUV promises to be valuable for large collaborative projects involving multiple laboratories, technicians, and/or sequencing platforms.
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              Variational Inference: A Review for Statisticians

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                Author and article information

                Contributors
                Role: Associate Editor
                Journal
                Bioinformatics
                Bioinformatics
                bioinformatics
                Bioinformatics
                Oxford University Press
                1367-4803
                1367-4811
                January 2024
                17 January 2024
                17 January 2024
                : 40
                : 1
                : btae024
                Affiliations
                Department of Statistics and Data Science, Southern Methodist University , Dallas, TX 75275, United States
                Department of Statistical Science, Baylor University , Waco, TX 76798, United States
                Department of Statistics and Data Science, Southern Methodist University , Dallas, TX 75275, United States
                Quantitative Biomedical Research Center, The University of Texas Southwestern Medical Center , Dallas, TX 75390, United States
                Department of Statistics and Data Science, Southern Methodist University , Dallas, TX 75275, United States
                Department of Mathematics, University of Texas at Arlington , Arlington, TX 76019 United States
                Division of Data Science, College of Science, University of Texas at Arlington , Arlington, TX 76019, United States
                Author notes
                Corresponding author. Department of Mathematics, University of Texas at Arlington, 411 S Nedderman Dr., Arlington, TX 76019, United States. E-mail: xinlei.wang@ 123456uta.edu (X.W.)
                Author information
                https://orcid.org/0000-0001-9387-9883
                https://orcid.org/0000-0002-8561-6511
                Article
                btae024
                10.1093/bioinformatics/btae024
                10826904
                38237909
                b552961a-e5f9-43b0-a7a9-56182f91b24c
                © The Author(s) 2024. Published by Oxford University Press.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 09 August 2023
                : 28 December 2023
                : 03 January 2024
                : 12 January 2024
                : 30 January 2024
                Page count
                Pages: 9
                Funding
                Funded by: NIGMS, DOI 10.13039/100000057;
                Award ID: R15GM131390
                Funded by: NCI, DOI 10.13039/100000054;
                Award ID: R01CA258584
                Funded by: NCI, DOI 10.13039/100000054;
                Award ID: U01CA249245
                Funded by: NIGMS, DOI 10.13039/100000057;
                Award ID: R01GM140012
                Funded by: NIGMS, DOI 10.13039/100000057;
                Award ID: R01GM141519
                Funded by: CPRIT, DOI 10.13039/100004917;
                Award ID: RP230330
                Categories
                Original Paper
                Gene Expression
                AcademicSubjects/SCI01060

                Bioinformatics & Computational biology
                Bioinformatics & Computational biology

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