Dataset Information

Divide-and-Conquer Matrisome Protein (DC-MaP) Strategy: An MS-Friendly Approach to Proteomic Matrisome Characterization.

ABSTRACT: Currently, the extracellular matrix (ECM) is considered a pivotal complex meshwork of macromolecules playing a plethora of biomolecular functions in health and disease beyond its commonly known mechanical role. Only by unraveling its composition can we leverage related tissue engineering and pharmacological efforts. Nevertheless, its unbiased proteomic identification still encounters some limitations mainly due to partial ECM enrichment by precipitation, sequential fractionation using unfriendly-mass spectrometry (MS) detergents, and resuspension with harsh reagents that need to be entirely removed prior to analysis. These methods can be technically challenging and labor-intensive, which affects the reproducibility of ECM identification and induces protein loss. Here, we present a simple new method applicable to tissue fragments of 10 mg and more. The technique has been validated on human ovarian tissue and involves a standardized procedure for sample processing with an MS-compatible detergent and combined centrifugation. This two-step protocol eliminates the need for laborious sample clarification and divides our samples into 2 fractions, soluble and insoluble, successively enriched with matrisome-associated (ECM-interacting) and core matrisome (structural ECM) proteins.

SUBMITTER: Ouni E

PROVIDER: S-EPMC7730167 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Publications

Divide-and-Conquer Matrisome Protein (DC-MaP) Strategy: An MS-Friendly Approach to Proteomic Matrisome Characterization.

Ouni Emna E Ruys Sebastien Pyr Dit SPD Dolmans Marie-Madeleine MM Herinckx Gaëtan G Vertommen Didier D Amorim Christiani A CA

International journal of molecular sciences 20201130 23

Currently, the extracellular matrix (ECM) is considered a pivotal complex meshwork of macromolecules playing a plethora of biomolecular functions in health and disease beyond its commonly known mechanical role. Only by unraveling its composition can we leverage related tissue engineering and pharmacological efforts. Nevertheless, its unbiased proteomic identification still encounters some limitations mainly due to partial ECM enrichment by precipitation, sequential fractionation using unfriendly ...[more]

PMID: 33266304

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Project description:Among proteins, orthologs are defined as those that are derived by vertical descent from a single progenitor in the last common ancestor of their host organisms. Our goal is to compute a complete set of protein orthologs derived from all currently available complete bacterial and archaeal genomes. Traditional approaches typically rely on all-against-all BLAST searching which is prohibitively expensive in terms of hardware requirements or computational time (requiring an estimated 18 months or more on a typical server). Here, we present xBASE-Orth, a system for ongoing ortholog annotation, which applies a "divide and conquer" approach and adopts a pragmatic scheme that trades accuracy for speed. Starting at species level, xBASE-Orth carefully constructs and uses pan-genomes as proxies for the full collections of coding sequences at each level as it progressively climbs the taxonomic tree using the previously computed data. This leads to a significant decrease in the number of alignments that need to be performed, which translates into faster computation, making ortholog computation possible on a global scale. Using xBASE-Orth, we analyzed an NCBI collection of 1,288 bacterial and 94 archaeal complete genomes with more than 4 million coding sequences in 5 weeks and predicted more than 700 million ortholog pairs, clustered in 175,531 orthologous groups. We have also identified sets of highly conserved bacterial and archaeal orthologs and in so doing have highlighted anomalies in genome annotation and in the proposed composition of the minimal bacterial genome. In summary, our approach allows for scalable and efficient computation of the bacterial and archaeal ortholog annotations. In addition, due to its hierarchical nature, it is suitable for incorporating novel complete genomes and alternative genome annotations. The computed ortholog data and a continuously evolving set of applications based on it are integrated in the xBASE database, available at http://www.xbase.ac.uk/.

Project description:Breast cancer claims 11,400 lives on average every year in the UK, making it one of the deadliest diseases. Mammography is the gold standard for detecting early signs of breast cancer, which can help cure the disease during its early stages. However, incorrect mammography diagnoses are common and may harm patients through unnecessary treatments and operations (or a lack of treatment). Therefore, systems that can learn to detect breast cancer on their own could help reduce the number of incorrect interpretations and missed cases. Various deep learning techniques, which can be used to implement a system that learns how to detect instances of breast cancer in mammograms, are explored throughout this paper. Convolution Neural Networks (CNNs) are used as part of a pipeline based on deep learning techniques. A divide and conquer approach is followed to analyse the effects on performance and efficiency when utilising diverse deep learning techniques such as varying network architectures (VGG19, ResNet50, InceptionV3, DenseNet121, MobileNetV2), class weights, input sizes, image ratios, pre-processing techniques, transfer learning, dropout rates, and types of mammogram projections. This approach serves as a starting point for model development of mammography classification tasks. Practitioners can benefit from this work by using the divide and conquer results to select the most suitable deep learning techniques for their case out-of-the-box, thus reducing the need for extensive exploratory experimentation. Multiple techniques are found to provide accuracy gains relative to a general baseline (VGG19 model using uncropped 512 × 512 pixels input images with a dropout rate of 0.2 and a learning rate of 1 × 10-3) on the Curated Breast Imaging Subset of DDSM (CBIS-DDSM) dataset. These techniques involve transfer learning pre-trained ImagetNet weights to a MobileNetV2 architecture, with pre-trained weights from a binarised version of the mini Mammography Image Analysis Society (mini-MIAS) dataset applied to the fully connected layers of the model, coupled with using weights to alleviate class imbalance, and splitting CBIS-DDSM samples between images of masses and calcifications. Using these techniques, a 5.6% gain in accuracy over the baseline model was accomplished. Other deep learning techniques from the divide and conquer approach, such as larger image sizes, do not yield increased accuracies without the use of image pre-processing techniques such as Gaussian filtering, histogram equalisation and input cropping.

Dataset Information

Divide-and-Conquer Matrisome Protein (DC-MaP) Strategy: An MS-Friendly Approach to Proteomic Matrisome Characterization.

Publications

Divide-and-Conquer Matrisome Protein (DC-MaP) Strategy: An MS-Friendly Approach to Proteomic Matrisome Characterization.

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