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Article ; Online: Treatment of advanced atherosclerotic mice with ABT-263 reduced indices of plaque stability and increased mortality.

Karnewar, Santosh / Karnewar, Vaishnavi / Shankman, Laura S / Owens, Gary K

JCI insight

2024 Volume 9, Issue 2

Abstract: The use of senolytic agents to remove senescent cells from atherosclerotic lesions is controversial. A common limitation of previous studies is the failure to rigorously define the effects of senolytic agent ABT-263 (Navitoclax) on smooth muscle cells ( ... ...

Abstract	The use of senolytic agents to remove senescent cells from atherosclerotic lesions is controversial. A common limitation of previous studies is the failure to rigorously define the effects of senolytic agent ABT-263 (Navitoclax) on smooth muscle cells (SMC) despite studies claiming that these cells are the major source of senescent cells. Moreover, there are no studies on the effect of ABT-263 on endothelial cells (EC), which - along with SMC - comprise 90% of α-smooth muscle actin+ (α-SMA+) myofibroblast-like cells in the protective fibrous cap. Here we tested the hypothesis that treatment of advanced atherosclerotic mice with ABT-263 will reduce lesion size and increase plaque stability. SMC (Myh11-CreERT2-eYFP) and EC (Cdh5-CreERT2-eYFP) lineage tracing Apoe-/- mice were fed a western diet (WD) for 18 weeks, followed by ABT-263 at 100 mg/kg/bw for 6 weeks or 50 mg/kg/bw for 9 weeks. ABT-263 treatment did not change lesion size or lumen area of the brachiocephalic artery (BCA). However, ABT-263 treatment reduced SMC by 90% and increased EC contributions to lesions via EC-to-mesenchymal transition (EndoMT) by 60%. ABT-263 treatment also reduced α-SMA+ fibrous cap thickness by 60% and was associated with a > 50% mortality rate. Taken together, ABT-263 treatment of WD-fed Apoe-/- mice with advanced lesions resulted in multiple detrimental changes, including reduced indices of stability and increased mortality.
MeSH term(s)	Mice ; Animals ; Endothelial Cells ; Mice, Knockout, ApoE ; Atherosclerosis/drug therapy ; Apolipoproteins E ; Aniline Compounds ; Sulfonamides
Chemical Substances	navitoclax (XKJ5VVK2WD) ; Apolipoproteins E ; Aniline Compounds ; Sulfonamides
Language	English
Publishing date	2024-01-23
Publishing country	United States
Document type	Journal Article
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.173863
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Treatment of advanced atherosclerotic mice with the senolytic agent ABT-263 is associated with reduced indices of plaque stability and increased mortality.

Karnewar, Santosh / Karnewar, Vaishnavi / Shankman, Laura S / Owens, Gary K

bioRxiv : the preprint server for biology

2023

Abstract	The use of senolytic agents to remove senescent cells from atherosclerotic lesions is controversial. A common limitation of previous studies is the failure to rigorously define the effects of senolytic agent ABT-263 (Navitoclax) on smooth muscle cells (SMC) despite studies claiming that they are the major source of senescent cells. Moreover, there are no studies of the effect of ABT-263 on endothelial cells (EC), which along with SMC comprise 90% of α-SMA
Language	English
Publishing date	2023-07-13
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.07.12.548696
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Book ; Online: Label-efficient Contrastive Learning-based model for nuclei detection and classification in 3D Cardiovascular Immunofluorescent Images

Moradinasab, Nazanin / Deaton, Rebecca A. / Shankman, Laura S. / Owens, Gary K. / Brown, Donald E.

2023

Abstract: Recently, deep learning-based methods achieved promising performance in nuclei detection and classification applications. However, training deep learning-based methods requires a large amount of pixel-wise annotated data, which is time-consuming and ... ...

Abstract	Recently, deep learning-based methods achieved promising performance in nuclei detection and classification applications. However, training deep learning-based methods requires a large amount of pixel-wise annotated data, which is time-consuming and labor-intensive, especially in 3D images. An alternative approach is to adapt weak-annotation methods, such as labeling each nucleus with a point, but this method does not extend from 2D histopathology images (for which it was originally developed) to 3D immunofluorescent images. The reason is that 3D images contain multiple channels (z-axis) for nuclei and different markers separately, which makes training using point annotations difficult. To address this challenge, we propose the Label-efficient Contrastive learning-based (LECL) model to detect and classify various types of nuclei in 3D immunofluorescent images. Previous methods use Maximum Intensity Projection (MIP) to convert immunofluorescent images with multiple slices to 2D images, which can cause signals from different z-stacks to falsely appear associated with each other. To overcome this, we devised an Extended Maximum Intensity Projection (EMIP) approach that addresses issues using MIP. Furthermore, we performed a Supervised Contrastive Learning (SCL) approach for weakly supervised settings. We conducted experiments on cardiovascular datasets and found that our proposed framework is effective and efficient in detecting and classifying various types of nuclei in 3D immunofluorescent images. Comment: 11 pages, 5 figures, MICCAI Workshop Conference 2023
Keywords	Electrical Engineering and Systems Science - Image and Video Processing ; Computer Science - Computer Vision and Pattern Recognition
Subject code	006
Publishing date	2023-09-07
Publishing country	us
Document type	Book ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: A New Autosomal

Deaton, Rebecca A / Bulut, Gamze / Serbulea, Vlad / Salamon, Anita / Shankman, Laura S / Nguyen, Anh Tram / Owens, Gary K

Arteriosclerosis, thrombosis, and vascular biology

2022 Volume 43, Issue 2, Page(s) 203–211

Abstract: Background: The : Methods: A : Results: Myh11-CreER: Conclusions: We generated and validated the function of an ... ...

Abstract	Background: The Methods: A Results: Myh11-CreER Conclusions: We generated and validated the function of an autosomal
MeSH term(s)	Mice ; Animals ; Male ; Female ; Mice, Transgenic ; Gene Knockout Techniques ; Integrases/genetics ; Integrases/metabolism ; Mice, Knockout ; Mice, Inbred C57BL ; Myocytes, Smooth Muscle/metabolism ; Cell Lineage ; Tamoxifen
Chemical Substances	Integrases (EC 2.7.7.-) ; Tamoxifen (094ZI81Y45)
Language	English
Publishing date	2022-12-15
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1221433-4
ISSN	1524-4636 ; 1079-5642
ISSN (online)	1524-4636
ISSN	1079-5642
DOI	10.1161/ATVBAHA.122.318160
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Article ; Online: IL-1β Inhibition Partially Negates the Beneficial Effects of Diet-Induced Atherosclerosis Regression in Mice.

Karnewar, Santosh / Karnewar, Vaishnavi / Deaton, Rebecca / Shankman, Laura S / Benavente, Ernest D / Williams, Corey M / Bradley, Xenia / Alencar, Gabriel F / Bulut, Gamze B / Kirmani, Sara / Baylis, Richard A / Zunder, Eli R / den Ruijter, Hester M / Pasterkamp, Gerard / Owens, Gary K

Arteriosclerosis, thrombosis, and vascular biology

2024

Abstract: Background: Thromboembolic events secondary to rupture or erosion of advanced atherosclerotic lesions is the global leading cause of death. The most common and effective means to reduce these major adverse cardiovascular events, including myocardial ... ...

Abstract	Background: Thromboembolic events secondary to rupture or erosion of advanced atherosclerotic lesions is the global leading cause of death. The most common and effective means to reduce these major adverse cardiovascular events, including myocardial infarction and stroke, is aggressive lipid lowering via a combination of drugs and dietary modifications. However, we know little regarding the effects of reducing dietary lipids on the composition and stability of advanced atherosclerotic lesions, the mechanisms that regulate these processes, and what therapeutic approaches might augment the benefits of lipid lowering. Methods: Smooth muscle cell lineage-tracing Results: Lipid lowering by switching Conclusions: Taken together, IL-1β appears to be required for the maintenance of standard laboratory diet-induced reductions in plaque burden and increases in multiple indices of plaque stability.
Language	English
Publishing date	2024-05-02
Publishing country	United States
Document type	Journal Article
ZDB-ID	1221433-4
ISSN	1524-4636 ; 1079-5642
ISSN (online)	1524-4636
ISSN	1079-5642
DOI	10.1161/ATVBAHA.124.320800
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Book ; Online: Weakly Supervised Deep Instance Nuclei Detection using Points Annotation in 3D Cardiovascular Immunofluorescent Images

Moradinasab, Nazanin / Sharma, Yash / Shankman, Laura S. / Owens, Gary K. / Brown, Donald E.

2022

Abstract: Two major causes of death in the United States and worldwide are stroke and myocardial infarction. The underlying cause of both is thrombi released from ruptured or eroded unstable atherosclerotic plaques that occlude vessels in the heart (myocardial ... ...

Abstract	Two major causes of death in the United States and worldwide are stroke and myocardial infarction. The underlying cause of both is thrombi released from ruptured or eroded unstable atherosclerotic plaques that occlude vessels in the heart (myocardial infarction) or the brain (stroke). Clinical studies show that plaque composition plays a more important role than lesion size in plaque rupture or erosion events. To determine the plaque composition, various cell types in 3D cardiovascular immunofluorescent images of plaque lesions are counted. However, counting these cells manually is expensive, time-consuming, and prone to human error. These challenges of manual counting motivate the need for an automated approach to localize and count the cells in images. The purpose of this study is to develop an automatic approach to accurately detect and count cells in 3D immunofluorescent images with minimal annotation effort. In this study, we used a weakly supervised learning approach to train the HoVer-Net segmentation model using point annotations to detect nuclei in fluorescent images. The advantage of using point annotations is that they require less effort as opposed to pixel-wise annotation. To train the HoVer-Net model using point annotations, we adopted a popularly used cluster labeling approach to transform point annotations into accurate binary masks of cell nuclei. Traditionally, these approaches have generated binary masks from point annotations, leaving a region around the object unlabeled (which is typically ignored during model training). However, these areas may contain important information that helps determine the boundary between cells. Therefore, we used the entropy minimization loss function in these areas to encourage the model to output more confident predictions on the unlabeled areas. Our comparison studies indicate that the HoVer-Net model trained using our weakly .
Keywords	Computer Science - Computer Vision and Pattern Recognition ; Computer Science - Artificial Intelligence
Subject code	006
Publishing date	2022-07-29
Publishing country	us
Document type	Book ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Efferocytosis by Paneth cells within the intestine.

Shankman, Laura S / Fleury, Samantha T / Evans, W Britt / Penberthy, Kristen K / Arandjelovic, Sanja / Blumberg, Richard S / Agaisse, Hervé / Ravichandran, Kodi S

Current biology : CB

2021 Volume 31, Issue 11, Page(s) 2469–2476.e5

Abstract: Apoptotic cells are quickly and efficiently engulfed and removed via the process of efferocytosis by either professional phagocytes, such as macrophages, or non-professional phagocytes, including epithelial cells. ...

Abstract	Apoptotic cells are quickly and efficiently engulfed and removed via the process of efferocytosis by either professional phagocytes, such as macrophages, or non-professional phagocytes, including epithelial cells.
MeSH term(s)	Animals ; Apoptosis ; Humans ; Inflammation ; Intestines ; Mice ; Paneth Cells ; Phagocytes ; Phagocytosis
Language	English
Publishing date	2021-04-13
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1071731-6
ISSN	1879-0445 ; 0960-9822
ISSN (online)	1879-0445
ISSN	0960-9822
DOI	10.1016/j.cub.2021.03.055
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Article: IL-1β inhibition partially negates the beneficial effects of diet-induced lipid lowering.

bioRxiv : the preprint server for biology

2023

Abstract: Background: Thromboembolic events secondary to rupture or erosion of advanced atherosclerotic lesions are the leading cause of death in the world. The most common and effective means to reduce these major adverse cardiovascular events (MACE), including ... ...

Abstract	Background: Thromboembolic events secondary to rupture or erosion of advanced atherosclerotic lesions are the leading cause of death in the world. The most common and effective means to reduce these major adverse cardiovascular events (MACE), including myocardial infarction (MI) and stroke, is aggressive lipid lowering via a combination of drugs and dietary modifications. However, little is known regarding the effects of reducing dietary lipids on the composition and stability of advanced atherosclerotic lesions, the mechanisms that regulate these processes, and what therapeutic approaches might augment the benefits of lipid lowering. Methods: Smooth muscle cell (SMC)-lineage tracing Results: Lipid-lowering by switching Conclusions: Taken together, IL-1β appears to be required for chow diet-induced reductions in plaque burden and increases in multiple indices of plaque stability.
Language	English
Publishing date	2023-10-14
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.10.13.562255
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Article ; Online: ELMO1 signaling is a promoter of osteoclast function and bone loss.

Arandjelovic, Sanja / Perry, Justin S A / Zhou, Ming / Ceroi, Adam / Smirnov, Igor / Walk, Scott F / Shankman, Laura S / Cambré, Isabelle / Onengut-Gumuscu, Suna / Elewaut, Dirk / Conrads, Thomas P / Ravichandran, Kodi S

Nature communications

2021 Volume 12, Issue 1, Page(s) 4974

Abstract: Osteoporosis affects millions worldwide and is often caused by osteoclast induced bone loss. Here, we identify the cytoplasmic protein ELMO1 as an important 'signaling node' in osteoclasts. We note that ELMO1 SNPs associate with bone abnormalities in ... ...

Abstract	Osteoporosis affects millions worldwide and is often caused by osteoclast induced bone loss. Here, we identify the cytoplasmic protein ELMO1 as an important 'signaling node' in osteoclasts. We note that ELMO1 SNPs associate with bone abnormalities in humans, and that ELMO1 deletion in mice reduces bone loss in four in vivo models: osteoprotegerin deficiency, ovariectomy, and two types of inflammatory arthritis. Our transcriptomic analyses coupled with CRISPR/Cas9 genetic deletion identify Elmo1 associated regulators of osteoclast function, including cathepsin G and myeloperoxidase. Further, we define the 'ELMO1 interactome' in osteoclasts via proteomics and reveal proteins required for bone degradation. ELMO1 also contributes to osteoclast sealing zone on bone-like surfaces and distribution of osteoclast-specific proteases. Finally, a 3D structure-based ELMO1 inhibitory peptide reduces bone resorption in wild type osteoclasts. Collectively, we identify ELMO1 as a signaling hub that regulates osteoclast function and bone loss, with relevance to osteoporosis and arthritis.
MeSH term(s)	Adaptor Proteins, Signal Transducing/deficiency ; Adaptor Proteins, Signal Transducing/genetics ; Adaptor Proteins, Signal Transducing/metabolism ; Animals ; Arthritis/pathology ; Bone Diseases, Metabolic/metabolism ; Bone Resorption/metabolism ; CRISPR-Cas Systems ; Female ; Mice ; Mice, Knockout ; Osteoclasts/metabolism ; Osteoporosis/metabolism ; Osteoprotegerin/deficiency ; Ovariectomy ; Signal Transduction ; Transcriptome ; X-Ray Microtomography
Chemical Substances	Adaptor Proteins, Signal Transducing ; ELMO1 protein, mouse ; Osteoprotegerin
Language	English
Publishing date	2021-08-17
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-021-25239-6
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Irradiation abolishes smooth muscle investment into vascular lesions in specific vascular beds.

Newman, Alexandra Ac / Baylis, Richard A / Hess, Daniel L / Griffith, Steven D / Shankman, Laura S / Cherepanova, Olga A / Owens, Gary K

JCI insight

2018 Volume 3, Issue 15

Abstract: The long-term adverse effects of radiotherapy on cardiovascular disease are well documented. However, the underlying mechanisms responsible for this increased risk are poorly understood. Previous studies using rigorous smooth muscle cell (SMC) lineage ... ...

Abstract	The long-term adverse effects of radiotherapy on cardiovascular disease are well documented. However, the underlying mechanisms responsible for this increased risk are poorly understood. Previous studies using rigorous smooth muscle cell (SMC) lineage tracing have shown abundant SMC investment into atherosclerotic lesions, where SMCs contribute to the formation of a protective fibrous cap. Studies herein tested whether radiation impairs protective adaptive SMC responses during vascular disease. To do this, we exposed SMC lineage tracing (Myh11-ERT2Cre YFP+) mice to lethal radiation (1,200 cGy) followed by bone marrow transplantation prior to atherosclerosis development or vessel injury. Surprisingly, following irradiation, we observed a complete loss of SMC investment in 100% of brachiocephalic artery (BCA), carotid artery, and aortic arch lesions. Importantly, this was associated with a decrease in multiple indices of atherosclerotic lesion stability within the BCA. Interestingly, we observed anatomic heterogeneity, as SMCs accumulated normally into lesions of the aortic root and abdominal aorta, suggesting that SMC sensitivity to lethal irradiation occurs in blood vessels of neural crest origin. Taken together, these results reveal an undefined and unintended variable in previous studies using lethal irradiation and may help explain why patients exposed to radiation have increased risk for cardiovascular disease.
MeSH term(s)	Animals ; Aorta, Abdominal/pathology ; Aorta, Abdominal/radiation effects ; Atherosclerosis/etiology ; Atherosclerosis/pathology ; Bone Marrow/radiation effects ; Bone Marrow Transplantation ; Brachiocephalic Trunk/pathology ; Brachiocephalic Trunk/radiation effects ; Cell Differentiation/radiation effects ; Disease Models, Animal ; Humans ; Male ; Mice ; Mice, Knockout, ApoE ; Muscle, Smooth, Vascular/cytology ; Muscle, Smooth, Vascular/radiation effects ; Myocytes, Smooth Muscle/radiation effects ; Whole-Body Irradiation
Language	English
Publishing date	2018-08-09
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.121017
Database	MEDical Literature Analysis and Retrieval System OnLINE

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