3de682901192d46ca6c10346a28e7bc2 pmc_34071374.pdf 021281f02cb44ed48a130a439a16f3fb6d6f20f7 pmc_34071374.pdf 745fab1e738f2e45951b6dd28ba50a508937e0f10ee58e714dce5cf260bf65c6 pmc_34071374.pdf Title: Unravelling the Encapsulation of DNA and Other Biomolecules in HAp Microcalcifications of Human Breast Cancer Tissues by Raman Imaging Subject: Microcalcifications are detected through mammography screening and, depending on their morphology and distribution (BI-RADS classification), they can be considered one of the first indicators of suspicious cancer lesions. However, the formation of hydroxyapatite (HAp) calcifications and their relationship with malignancy remains unknown. In this work, we report the most detailed three-dimensional biochemical analysis of breast cancer microcalcifications to date, combining 3D Raman spectroscopy imaging and advanced multivariate analysis in order to investigate in depth the molecular composition of HAp calcifications found in 26 breast cancer tissue biopsies. We demonstrate that DNA has been naturally adsorbed and encapsulated inside HAp microcalcifications. Furthermore, we also show the encapsulation of other relevant biomolecules in HAp calcifications, such as lipids, proteins, cytochrome C and polysaccharides. The demonstration of natural DNA biomineralization, particularly in the tumor microenvironment, represents an unprecedented advance in the field, as it can pave the way to understanding the role of HAp in malignant tissues. Keywords: microcalcifications; DNA mineralization; breast cancer; tumor microenvironment; Raman spectroscopy Author: Monica Marro, Anna Maria Rodríguez-Rivero, Cuauhtémoc Araujo-Andrade, Maria Teresa Fernández-Figueras, Laia Pérez-Roca, Eva Castellà, Jordi Navinés, Antonio Mariscal, Joan Francesc Julián, Pau Turon and Pablo Loza-Alvarez Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Jun 1 09:07:38 2021 CEST ModDate: Tue Jun 1 09:16:14 2021 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3602917 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SXQPWE+VnURWPalladioL Type 1 Custom yes yes no 10 0 ZXKVHA+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 KHKWPJ+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 JXJOWY+URWPalladioL-Roma Type 1 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FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 167 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 173 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 179 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 188 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 194 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 200 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 215 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 221 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 227 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-10-21 02:37:48 CEST RepresentationInformation: pmc_34071374.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-03-31 12:14:04 CEST Size: 3602917 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 469 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Children: Item: Title: Breast Tumor Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Raman Spectroscopy Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Multivariate Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Breast Tumor Sample Staining for Confocal Microscopy Imaging Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: In Vitro Synthesis of DNA-HAp Particles Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Children: Item: Title: Molecular Raman Imaging and Advanced Multivariate Analysis of Tissue Calcifications Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Study of DNA Mineralization in HAp Microcalcifications Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Children: Item: Title: Reference Raman Spectral Databases and Literature Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Identification of DNA in HAp Calcifications Found in Breast Cancer Tissues Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Fluorescence Staining of Microcalcifications with DAPI Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Raman Images Reveal the Presence and Distribution of Biomolecules Inside HAp Calcifications Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Info: Title: Unravelling the Encapsulation of DNA and Other Biomolecules in HAp Microcalcifications of Human Breast Cancer Tissues by Raman Imaging Author: Monica Marro, Anna Maria Rodríguez-Rivero, Cuauhtémoc Araujo-Andrade, Maria Teresa Fernández-Figueras, Laia Pérez-Roca, Eva Castellà, Jordi Navinés, Antonio Mariscal, Joan Francesc Julián, Pau Turon and Pablo Loza-Alvarez Subject: Microcalcifications are detected through mammography screening and, depending on their morphology and distribution (BI-RADS classification), they can be considered one of the first indicators of suspicious cancer lesions. However, the formation of hydroxyapatite (HAp) calcifications and their relationship with malignancy remains unknown. In this work, we report the most detailed three-dimensional biochemical analysis of breast cancer microcalcifications to date, combining 3D Raman spectroscopy imaging and advanced multivariate analysis in order to investigate in depth the molecular composition of HAp calcifications found in 26 breast cancer tissue biopsies. We demonstrate that DNA has been naturally adsorbed and encapsulated inside HAp microcalcifications. Furthermore, we also show the encapsulation of other relevant biomolecules in HAp calcifications, such as lipids, proteins, cytochrome C and polysaccharides. The demonstration of natural DNA biomineralization, particularly in the tumor microenvironment, represents an unprecedented advance in the field, as it can pave the way to understanding the role of HAp in malignant tissues. 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