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Article ; Online: Author Correction: Automated segmentation and tracking of mitochondria in live-cell time-lapse images.

Lefebvre, Austin E Y T / Ma, Dennis / Kessenbrock, Kai / Lawson, Devon A / Digman, Michelle A

2022 Volume 19, Issue 6, Page(s) 770

Language	English
Publishing date	2022-04-29
Publishing country	United States
Document type	Published Erratum
ZDB-ID	2169522-2
ISSN	1548-7105 ; 1548-7091
ISSN (online)	1548-7105
ISSN	1548-7091
DOI	10.1038/s41592-022-01506-2
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Article ; Online: Automated segmentation and tracking of mitochondria in live-cell time-lapse images.

Lefebvre, Austin E Y T / Ma, Dennis / Kessenbrock, Kai / Lawson, Devon A / Digman, Michelle A

Nature methods

2021 Volume 18, Issue 9, Page(s) 1091–1102

Abstract: Mitochondria display complex morphology and movements, which complicates their segmentation and tracking in time-lapse images. Here, we introduce Mitometer, an algorithm for fast, unbiased, and automated segmentation and tracking of mitochondria in live- ... ...

Abstract	Mitochondria display complex morphology and movements, which complicates their segmentation and tracking in time-lapse images. Here, we introduce Mitometer, an algorithm for fast, unbiased, and automated segmentation and tracking of mitochondria in live-cell two-dimensional and three-dimensional time-lapse images. Mitometer requires only the pixel size and the time between frames to identify mitochondrial motion and morphology, including fusion and fission events. The segmentation algorithm isolates individual mitochondria via a shape- and size-preserving background removal process. The tracking algorithm links mitochondria via differences in morphological features and displacement, followed by a gap-closing scheme. Using Mitometer, we show that mitochondria of triple-negative breast cancer cells are faster, more directional, and more elongated than those in their receptor-positive counterparts. Furthermore, we show that mitochondrial motility and morphology in breast cancer, but not in normal breast epithelia, correlate with metabolic activity. Mitometer is an unbiased and user-friendly tool that will help resolve fundamental questions regarding mitochondrial form and function.
MeSH term(s)	Algorithms ; Breast Neoplasms/metabolism ; Breast Neoplasms/pathology ; Cells, Cultured ; Female ; Humans ; Imaging, Three-Dimensional/methods ; Mammary Glands, Human/cytology ; Mitochondria/metabolism ; NAD/metabolism ; Reproducibility of Results ; Software ; Time-Lapse Imaging/methods ; Triple Negative Breast Neoplasms/pathology
Chemical Substances	NAD (0U46U6E8UK)
Language	English
Publishing date	2021-08-19
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2169522-2
ISSN	1548-7105 ; 1548-7091
ISSN (online)	1548-7105
ISSN	1548-7091
DOI	10.1038/s41592-021-01234-z
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Tumour heterogeneity and metastasis at single-cell resolution.

Lawson, Devon A / Kessenbrock, Kai / Davis, Ryan T / Pervolarakis, Nicholas / Werb, Zena

Nature cell biology

2018 Volume 20, Issue 12, Page(s) 1349–1360

Abstract: Tumours comprise a heterogeneous collection of cells with distinct genetic and phenotypic properties that can differentially promote progression, metastasis and drug resistance. Emerging single-cell technologies provide a new opportunity to profile ... ...

Abstract	Tumours comprise a heterogeneous collection of cells with distinct genetic and phenotypic properties that can differentially promote progression, metastasis and drug resistance. Emerging single-cell technologies provide a new opportunity to profile individual cells within tumours and investigate what roles they play in these processes. This Review discusses key technological considerations for single-cell studies in cancer, new findings using single-cell technologies and critical open questions for future applications.
MeSH term(s)	Biomarkers, Tumor/genetics ; Biomarkers, Tumor/metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Heterogeneity ; Humans ; Neoplasm Metastasis ; Neoplasms/genetics ; Neoplasms/metabolism ; Neoplasms/pathology ; Neoplastic Cells, Circulating/metabolism ; Neoplastic Cells, Circulating/pathology ; Single-Cell Analysis/methods ; Tumor Microenvironment/genetics
Chemical Substances	Biomarkers, Tumor
Language	English
Publishing date	2018-11-26
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1474722-4
ISSN	1476-4679 ; 1465-7392
ISSN (online)	1476-4679
ISSN	1465-7392
DOI	10.1038/s41556-018-0236-7
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Article ; Online: The Cleared Mammary Fat Pad Transplantation Assay for Mammary Epithelial Organogenesis.

Lawson, Devon A / Werb, Zena / Zong, Yang / Goldstein, Andrew S

Cold Spring Harbor protocols

2015 Volume 2015, Issue 12, Page(s) pdb.prot078071

Abstract: Cleared mammary fat pad (MFP) transplantation has been a standard technique for studies of mammary development and cancer for several decades. The mammary gland is comprised of several fundamental components: The epithelial compartment contains basal/ ... ...

Abstract	Cleared mammary fat pad (MFP) transplantation has been a standard technique for studies of mammary development and cancer for several decades. The mammary gland is comprised of several fundamental components: The epithelial compartment contains basal/myoepithelial cells and luminal cells, and the stromal compartment (called the MFP) contains adipocytes, smooth muscle cells, fibroblasts, and immune cells. In 3- to 4-wk-old female mice, the mammary epithelium is concentrated very close to the nipple and has not yet grown beyond the mammary lymph node to penetrate the bulk of the MFP. This developmental feature provides an anatomical fixed point, and enables one to cut away the portion of the MFP from the nipple to the lymph node, leaving behind the majority of the MFP free of epithelium. The "cleared" MFP can serve as a supportive native microenvironment fully sufficient for the organogenesis of injected donor epithelium. Normal mammary epithelial donor cells will produce histologically and functionally normal mammary ductal epithelium several weeks posttransplant, with the exception that the ducts will not be connected to the nipple. The assay described here provides a powerful platform for assessing the developmental and tumorigenic potential of engineered cells of interest.
MeSH term(s)	Adipose Tissue/anatomy & histology ; Adipose Tissue/physiology ; Animals ; Carcinogenesis ; Epithelial Cells/physiology ; Humans ; Mammary Glands, Human/anatomy & histology ; Mammary Glands, Human/growth & development ; Mice ; Models, Animal ; Organogenesis ; Transplantation/methods
Language	English
Publishing date	2015-12-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	1559-6095
ISSN (online)	1559-6095
DOI	10.1101/pdb.prot078071
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Article ; Online: Lattice light sheet imaging of membrane nanotubes between human breast cancer cells in culture and in brain metastases.

Parker, Ian / Evans, Katrina T / Ellefsen, Kyle / Lawson, Devon A / Smith, Ian F

Scientific reports

2017 Volume 7, Issue 1, Page(s) 11029

Abstract: Membrane nanotubes are cytosolic protrusions with diameters <1 µm that extend between cells separated by tens of µm. They mediate several forms of intercellular communication and are upregulated in diverse diseases. Difficulties in visualizing and ... ...

Abstract	Membrane nanotubes are cytosolic protrusions with diameters <1 µm that extend between cells separated by tens of µm. They mediate several forms of intercellular communication and are upregulated in diverse diseases. Difficulties in visualizing and studying nanotubes within intact tissues have, however, prompted skepticism regarding their in vivo relevance, and most studies have been confined to cell culture systems. Here, we introduce lattice-light sheet imaging of MDA-MB-231 human breast cancer cells genetically engineered to brightly express membrane-targeted GFP as a promising approach to visualize membrane nanotubes in vitro and in situ. We demonstrate that cultured cells form multiple nanotubes that mediate intercellular communication of Ca
MeSH term(s)	Animals ; Brain/pathology ; Brain Neoplasms/pathology ; Brain Neoplasms/secondary ; Breast Neoplasms/pathology ; Cell Culture Techniques ; Cell Surface Extensions/ultrastructure ; Disease Models, Animal ; Genes, Reporter ; Green Fluorescent Proteins/analysis ; Humans ; Mice ; Neoplasm Metastasis/pathology ; Staining and Labeling/methods ; Tumor Cells, Cultured/ultrastructure
Chemical Substances	Green Fluorescent Proteins (147336-22-9)
Language	English
Publishing date	2017-09-08
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-017-11223-y
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Microglia promote anti-tumour immunity and suppress breast cancer brain metastasis.

Evans, Katrina T / Blake, Kerrigan / Longworth, Aaron / Coburn, Morgan A / Insua-Rodríguez, Jacob / McMullen, Timothy P / Nguyen, Quy H / Ma, Dennis / Lev, Tatyana / Hernandez, Grace A / Oganyan, Armani K / Orujyan, Davit / Edwards, Robert A / Pridans, Clare / Green, Kim N / Villalta, S Armando / Blurton-Jones, Mathew / Lawson, Devon A

Nature cell biology

2023 Volume 25, Issue 12, Page(s) 1848–1859

Abstract: Breast cancer brain metastasis (BCBM) is a lethal disease with no effective treatments. Prior work has shown that brain cancers and metastases are densely infiltrated with anti-inflammatory, protumourigenic tumour-associated macrophages, but the role of ... ...

Abstract	Breast cancer brain metastasis (BCBM) is a lethal disease with no effective treatments. Prior work has shown that brain cancers and metastases are densely infiltrated with anti-inflammatory, protumourigenic tumour-associated macrophages, but the role of brain-resident microglia remains controversial because they are challenging to discriminate from other tumour-associated macrophages. Using single-cell RNA sequencing, genetic and humanized mouse models, we specifically identify microglia and find that they play a distinct pro-inflammatory and tumour-suppressive role in BCBM. Animals lacking microglia show increased metastasis, decreased survival and reduced natural killer and T cell responses, showing that microglia are critical to promote anti-tumour immunity to suppress BCBM. We find that the pro-inflammatory response is conserved in human microglia, and markers of their response are associated with better prognosis in patients with BCBM. These findings establish an important role for microglia in anti-tumour immunity and highlight them as a potential immunotherapy target for brain metastasis.
MeSH term(s)	Mice ; Animals ; Humans ; Female ; Microglia ; Breast Neoplasms/genetics ; Breast Neoplasms/pathology ; Brain Neoplasms/pathology ; Brain/pathology ; Treatment Outcome
Language	English
Publishing date	2023-11-13
Publishing country	England
Document type	Journal Article
ZDB-ID	1474722-4
ISSN	1476-4679 ; 1465-7392
ISSN (online)	1476-4679
ISSN	1465-7392
DOI	10.1038/s41556-023-01273-y
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Article: Stem cells in prostate cancer initiation and progression.

Lawson, Devon A / Witte, Owen N

The Journal of clinical investigation

2007 Volume 117, Issue 8, Page(s) 2044–2050

Abstract: Peter Nowell and David Hungerford's discovery of the Philadelphia chromosome facilitated many critical studies that have led to a paradigm shift in our understanding of cancer as a disease of stem cells. This Review focuses on the application of these ... ...

Abstract	Peter Nowell and David Hungerford's discovery of the Philadelphia chromosome facilitated many critical studies that have led to a paradigm shift in our understanding of cancer as a disease of stem cells. This Review focuses on the application of these concepts to investigation of the role of stem cells in prostate cancer initiation and progression. Major strides in the development of in vitro and in vivo assays have enabled identification and characterization of prostate stem cells as well as functional evaluation of the tumorigenic effects of prostate cancer-related genetic alterations.
MeSH term(s)	Animals ; Cell Transformation, Neoplastic ; Humans ; Male ; Neoplastic Stem Cells/metabolism ; Neoplastic Stem Cells/pathology ; Philadelphia Chromosome ; Prostatic Neoplasms/genetics ; Prostatic Neoplasms/metabolism ; Prostatic Neoplasms/pathology ; Stem Cells/metabolism ; Stem Cells/pathology
Language	English
Publishing date	2007-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	3067-3
ISSN	1558-8238 ; 0021-9738
ISSN (online)	1558-8238
ISSN	0021-9738
DOI	10.1172/JCI32810
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Patient-derived xenograft culture-transplant system for investigation of human breast cancer metastasis.

Ma, Dennis / Hernandez, Grace A / Lefebvre, Austin E Y T / Alshetaiwi, Hamad / Blake, Kerrigan / Dave, Kushal R / Rauf, Maha / Williams, Justice W / Davis, Ryan T / Evans, Katrina T / Longworth, Aaron / Masoud, Madona Y G / Lee, Regis / Edwards, Robert A / Digman, Michelle A / Kessenbrock, Kai / Lawson, Devon A

Communications biology

2021 Volume 4, Issue 1, Page(s) 1268

Abstract: Metastasis is a fatal disease where research progress has been hindered by a lack of authentic experimental models. Here, we develop a 3D tumor sphere culture-transplant system that facilitates the growth and engineering of patient-derived xenograft (PDX) ...

Abstract	Metastasis is a fatal disease where research progress has been hindered by a lack of authentic experimental models. Here, we develop a 3D tumor sphere culture-transplant system that facilitates the growth and engineering of patient-derived xenograft (PDX) tumor cells for functional metastasis assays in vivo. Orthotopic transplantation and RNA sequencing (RNA-seq) analyses show that PDX tumor spheres maintain tumorigenic potential, and the molecular marker and global transcriptome signatures of native tumor cells. Tumor spheres display robust capacity for lentiviral engineering and dissemination in spontaneous and experimental metastasis assays in vivo. Inhibition of pathways previously reported to attenuate metastasis also inhibit metastasis after sphere culture, validating our approach for authentic investigations of metastasis. Finally, we demonstrate a new role for the metabolic enzyme NME1 in promoting breast cancer metastasis, providing proof-of-principle that our culture-transplant system can be used for authentic propagation and engineering of patient tumor cells for functional studies of metastasis.
MeSH term(s)	Animals ; Breast Neoplasms/pathology ; Disease Models, Animal ; Female ; Heterografts ; Mice ; Neoplasm Metastasis ; Neoplasms, Experimental ; Tumor Microenvironment ; Xenograft Model Antitumor Assays
Language	English
Publishing date	2021-11-05
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	2399-3642
ISSN (online)	2399-3642
DOI	10.1038/s42003-021-02596-y
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: A spatially resolved single cell genomic atlas of the adult human breast.

Kumar, Tapsi / Nee, Kevin / Wei, Runmin / He, Siyuan / Nguyen, Quy H / Bai, Shanshan / Blake, Kerrigan / Gong, Yanwen / Pein, Maren / Sei, Emi / Hu, Min / Casasent, Anna / Thennavan, Aatish / Li, Jianzhuo / Tran, Tuan / Chen, Ken / Nilges, Benedikt / Kashikar, Nachiket / Braubach, Oliver /

Cheikh, Bassem Ben / Nikulina, Nadya / Chen, Hui / Teshome, Mediget / Menegaz, Brian / Javaid, Huma / Nagi, Chandandeep / Montalvan, Jessica / Tifrea, Delia F / Edwards, Robert / Lin, Erin / Parajuli, Ritesh / Winocour, Sebastian / Thompson, Alastair / Lim, Bora / Lawson, Devon A / Kessenbrock, Kai / Navin, Nicholas

bioRxiv : the preprint server for biology

2023

Abstract: The adult human breast comprises an intricate network of epithelial ducts and lobules that are embedded in connective and adipose tissue. While previous studies have mainly focused on the breast epithelial system, many of the non-epithelial cell types ... ...

Abstract	The adult human breast comprises an intricate network of epithelial ducts and lobules that are embedded in connective and adipose tissue. While previous studies have mainly focused on the breast epithelial system, many of the non-epithelial cell types remain understudied. Here, we constructed a comprehensive Human Breast Cell Atlas (HBCA) at single-cell and spatial resolution. Our single-cell transcriptomics data profiled 535,941 cells from 62 women, and 120,024 nuclei from 20 women, identifying 11 major cell types and 53 cell states. These data revealed abundant pericyte, endothelial and immune cell populations, and highly diverse luminal epithelial cell states. Our spatial mapping using three technologies revealed an unexpectedly rich ecosystem of tissue-resident immune cells in the ducts and lobules, as well as distinct molecular differences between ductal and lobular regions. Collectively, these data provide an unprecedented reference of adult normal breast tissue for studying mammary biology and disease states such as breast cancer.
Language	English
Publishing date	2023-04-25
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.04.22.537946
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Article ; Online: Transcriptional diversity and bioenergetic shift in human breast cancer metastasis revealed by single-cell RNA sequencing.

Davis, Ryan T / Blake, Kerrigan / Ma, Dennis / Gabra, Mari B Ishak / Hernandez, Grace A / Phung, Anh T / Yang, Ying / Maurer, Dustin / Lefebvre, Austin E Y T / Alshetaiwi, Hamad / Xiao, Zhengtao / Liu, Juan / Locasale, Jason W / Digman, Michelle A / Mjolsness, Eric / Kong, Mei / Werb, Zena / Lawson, Devon A

Nature cell biology

2020 Volume 22, Issue 3, Page(s) 310–320

Abstract: Although metastasis remains the cause of most cancer-related mortality, mechanisms governing seeding in distal tissues are poorly understood. Here, we establish a robust method for the identification of global transcriptomic changes in rare metastatic ... ...

Abstract	Although metastasis remains the cause of most cancer-related mortality, mechanisms governing seeding in distal tissues are poorly understood. Here, we establish a robust method for the identification of global transcriptomic changes in rare metastatic cells during seeding using single-cell RNA sequencing and patient-derived-xenograft models of breast cancer. We find that both primary tumours and micrometastases display transcriptional heterogeneity but micrometastases harbour a distinct transcriptome program conserved across patient-derived-xenograft models that is highly predictive of poor survival of patients. Pathway analysis revealed mitochondrial oxidative phosphorylation as the top pathway upregulated in micrometastases, in contrast to higher levels of glycolytic enzymes in primary tumour cells, which we corroborated by flow cytometric and metabolomic analyses. Pharmacological inhibition of oxidative phosphorylation dramatically attenuated metastatic seeding in the lungs, which demonstrates the functional importance of oxidative phosphorylation in metastasis and highlights its potential as a therapeutic target to prevent metastatic spread in patients with breast cancer.
MeSH term(s)	Animals ; Breast Neoplasms/genetics ; Breast Neoplasms/metabolism ; Breast Neoplasms/pathology ; Energy Metabolism ; Female ; Humans ; Mice, Inbred NOD ; Mice, SCID ; Mitochondria/metabolism ; Neoplasm Metastasis ; Oxidative Phosphorylation ; Sequence Analysis, RNA ; Single-Cell Analysis ; Transcription, Genetic ; Transcriptome
Language	English
Publishing date	2020-03-06
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	1474722-4
ISSN	1476-4679 ; 1465-7392
ISSN (online)	1476-4679
ISSN	1465-7392
DOI	10.1038/s41556-020-0477-0
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