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Article ; Online: Cytoplasmic retention of the DNA/RNA-binding protein FUS ameliorates organ fibrosis in mice.

Chiusa, Manuel / Lee, Youngmin A / Zhang, Ming-Zhi / Harris, Raymond C / Sherrill, Taylor / Lindner, Volkhard / Brooks, Craig R / Yu, Gang / Fogo, Agnes B / Flynn, Charles R / Zienkiewicz, Jozef / Hawiger, Jacek / Zent, Roy / Pozzi, Ambra

The Journal of clinical investigation

2024 Volume 134, Issue 6

Abstract: Uncontrolled accumulation of extracellular matrix leads to tissue fibrosis and loss of organ function. We previously demonstrated in vitro that the DNA/RNA-binding protein fused in sarcoma (FUS) promotes fibrotic responses by translocating to the nucleus, ...

Abstract	Uncontrolled accumulation of extracellular matrix leads to tissue fibrosis and loss of organ function. We previously demonstrated in vitro that the DNA/RNA-binding protein fused in sarcoma (FUS) promotes fibrotic responses by translocating to the nucleus, where it initiates collagen gene transcription. However, it is still not known whether FUS is profibrotic in vivo and whether preventing its nuclear translocation might inhibit development of fibrosis following injury. We now demonstrate that levels of nuclear FUS are significantly increased in mouse models of kidney and liver fibrosis. To evaluate the direct role of FUS nuclear translocation in fibrosis, we used mice that carry a mutation in the FUS nuclear localization sequence (FUSR521G) and the cell-penetrating peptide CP-FUS-NLS that we previously showed inhibits FUS nuclear translocation in vitro. We provide evidence that FUSR521G mice or CP-FUS-NLS-treated mice showed reduced nuclear FUS and fibrosis following injury. Finally, differential gene expression analysis and immunohistochemistry of tissues from individuals with focal segmental glomerulosclerosis or nonalcoholic steatohepatitis revealed significant upregulation of FUS and/or collagen genes and FUS protein nuclear localization in diseased organs. These results demonstrate that injury-induced nuclear translocation of FUS contributes to fibrosis and highlight CP-FUS-NLS as a promising therapeutic option for organ fibrosis.
MeSH term(s)	Animals ; Mice ; RNA ; RNA-Binding Protein FUS/genetics ; RNA-Binding Protein FUS/metabolism ; DNA-Binding Proteins/genetics ; RNA-Binding Proteins/genetics ; RNA-Binding Proteins/metabolism ; Mutation ; DNA ; Fibrosis ; Collagen/metabolism ; Amyotrophic Lateral Sclerosis/genetics
Chemical Substances	RNA (63231-63-0) ; RNA-Binding Protein FUS ; DNA-Binding Proteins ; RNA-Binding Proteins ; DNA (9007-49-2) ; Collagen (9007-34-5)
Language	English
Publishing date	2024-03-15
Publishing country	United States
Document type	Journal Article
ZDB-ID	3067-3
ISSN	1558-8238 ; 0021-9738
ISSN (online)	1558-8238
ISSN	0021-9738
DOI	10.1172/JCI175158
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Article ; Online: Perfusion and Inflation of the Mouse Lung for Tumor Histology.

Davenport, Mackenzie L / Sherrill, Taylor P / Blackwell, Timothy S / Edmonds, Mick D

Journal of visualized experiments : JoVE

2020 , Issue 162

Abstract: The ability to evaluate lung histology is critical for the fields of lung cancer research and cancer metastasis. It is equally important to perform necropsies rapidly and efficiently from studies without sacrificing the quality of the tissues procured. ... ...

Abstract	The ability to evaluate lung histology is critical for the fields of lung cancer research and cancer metastasis. It is equally important to perform necropsies rapidly and efficiently from studies without sacrificing the quality of the tissues procured. The goal of this protocol is to present a method to rapidly perfuse, inflate, and fix mouse lungs for downstream histological analysis. This method does not standardize lung inflation; thus, it does not require any special procedures or equipment and instead simply instills fixative directly through the trachea following perfusion through the heart. This allows for sufficient estimation of tumor size, histology, and scoring. This also allows for the collection of frozen tissue prior to lung tissue fixation. This method is limited in that it does not allow for later morphometric quantification of the lung; however, it is more than sufficient for lung tumor analysis from genetically engineered mouse models (GEMMs), syngeneic models, as well as xenograft tumor and metastasis studies.
MeSH term(s)	Animals ; Humans ; Lung/pathology ; Lung Neoplasms/pathology ; Mice ; Perfusion ; Staining and Labeling ; Tissue Fixation ; Xenograft Model Antitumor Assays
Language	English
Publishing date	2020-08-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/60605
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Article: Perfusion and inflation of the mouse lung for tumor histology

Davenport, Mackenzie L / Sherrill, Taylor P / Blackwell, Timothy S / Edmonds, Mick D

Journal of visualized experiments. 2020 Aug. 06, , no. 162

2020

Abstract	The ability to evaluate lung histology is critical for the fields of lung cancer research and cancer metastasis. It is equally important to perform necropsies rapidly and efficiently from studies without sacrificing the quality of the tissues procured. The goal of this protocol is to present a method to rapidly perfuse, inflate, and fix mouse lungs for downstream histological analysis. This method does not standardize lung inflation; thus, it does not require any special procedures or equipment and instead simply instills fixative directly through the trachea following perfusion through the heart. This allows for sufficient estimation of tumor size, histology, and scoring. This also allows for the collection of frozen tissue prior to lung tissue fixation. This method is limited in that it does not allow for later morphometric quantification of the lung; however, it is more than sufficient for lung tumor analysis from genetically engineered mouse models (GEMMs), syngeneic models, as well as xenograft tumor and metastasis studies.
Keywords	equipment ; genetic engineering ; heart ; histology ; inflation ; lung neoplasms ; lungs ; metastasis ; mice ; morphometry ; xenotransplantation
Language	English
Dates of publication	2020-0806
Size	p. e60605.
Publishing place	Journal of Visualized Experiments
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/60605
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Article ; Online: Enhanced Expression of Catalase in Mitochondria Modulates NF-κB-Dependent Lung Inflammation through Alteration of Metabolic Activity in Macrophages.

Han, Wei / Fessel, Joshua P / Sherrill, Taylor / Kocurek, Emily G / Yull, Fiona E / Blackwell, Timothy S

Journal of immunology (Baltimore, Md. : 1950)

2020 Volume 205, Issue 4, Page(s) 1125–1134

Abstract: NF-κB is a reduction-oxidation-sensitive transcription factor that plays a key role in regulating the immune response. In these studies, we intended to investigate the role of mitochondrial-derived reactive oxygen species in regulating NF-κB activation ... ...

Abstract	NF-κB is a reduction-oxidation-sensitive transcription factor that plays a key role in regulating the immune response. In these studies, we intended to investigate the role of mitochondrial-derived reactive oxygen species in regulating NF-κB activation by studying transgenic mice that overexpress mitochondrial-targeted human catalase (mCAT). We treated wild-type (WT) and mCAT mice with intratracheal instillation of
MeSH term(s)	Animals ; Bone Marrow/metabolism ; Catalase/metabolism ; Lung/metabolism ; Macrophages/metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Mitochondria/metabolism ; NAD/metabolism ; NF-kappa B/metabolism ; Pneumonia/metabolism ; Reactive Oxygen Species/metabolism ; Signal Transduction/physiology
Chemical Substances	NF-kappa B ; Reactive Oxygen Species ; NAD (0U46U6E8UK) ; Catalase (EC 1.11.1.6)
Language	English
Publishing date	2020-06-29
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	3056-9
ISSN	1550-6606 ; 0022-1767 ; 1048-3233 ; 1047-7381
ISSN (online)	1550-6606
ISSN	0022-1767 ; 1048-3233 ; 1047-7381
DOI	10.4049/jimmunol.1900820
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Article ; Online: Secretory Cells Are the Primary Source of pIgR in Small Airways.

Blackburn, Jessica B / Schaff, Jacob A / Gutor, Sergey / Du, Rui-Hong / Nichols, David / Sherrill, Taylor / Gutierrez, Austin J / Xin, Matthew K / Wickersham, Nancy / Zhang, Yong / Holtzman, Michael J / Ware, Lorraine B / Banovich, Nicholas E / Kropski, Jonathan A / Blackwell, Timothy S / Richmond, Bradley W

American journal of respiratory cell and molecular biology

2022 Volume 67, Issue 3, Page(s) 334–345

Abstract: Loss of secretory IgA (SIgA) is common in chronic obstructive pulmonary disease (COPD) small airways and likely contributes to disease progression. We hypothesized that loss of SIgA results from reduced expression of pIgR (polymeric immunoglobulin ... ...

Abstract	Loss of secretory IgA (SIgA) is common in chronic obstructive pulmonary disease (COPD) small airways and likely contributes to disease progression. We hypothesized that loss of SIgA results from reduced expression of pIgR (polymeric immunoglobulin receptor), a chaperone protein needed for SIgA transcytosis, in the COPD small airway epithelium. pIgR-expressing cells were defined and quantified at single-cell resolution in human airways using RNA
MeSH term(s)	Animals ; Haemophilus influenzae/enzymology ; Humans ; Immunoglobulin A, Secretory/metabolism ; Leukocyte Elastase/metabolism ; Mice ; Proteolysis ; Pulmonary Disease, Chronic Obstructive/metabolism ; Receptors, Polymeric Immunoglobulin/genetics ; Receptors, Polymeric Immunoglobulin/metabolism ; Respiratory System/metabolism
Chemical Substances	Immunoglobulin A, Secretory ; Receptors, Polymeric Immunoglobulin ; Leukocyte Elastase (EC 3.4.21.37)
Language	English
Publishing date	2022-02-17
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	1025960-0
ISSN	1535-4989 ; 1044-1549
ISSN (online)	1535-4989
ISSN	1044-1549
DOI	10.1165/rcmb.2021-0548OC
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Article: Novel mouse model of indwelling pleural catheter in mice with malignant pleural effusion.

Merrick, Christopher / Sherrill, Taylor / Kanellakis, Nikolaos I / Asciak, Rachelle / Stathopoulos, Georgios T / Maldonado, Fabien / Rahman, Najib M / Blackwell, Timothy / Psallidas, Ioannis

ERJ open research

2019 Volume 5, Issue 2

Abstract: This novel mouse model mimics malignant pleural effusion drainage using an indwelling pleural catheter in humans, and provides direct access to the pleural space potentially enabling the testing of intrapleural therapies in the treatment of MPE. ...

Abstract	This novel mouse model mimics malignant pleural effusion drainage using an indwelling pleural catheter in humans, and provides direct access to the pleural space potentially enabling the testing of intrapleural therapies in the treatment of MPE.
Language	English
Publishing date	2019-05-28
Publishing country	England
Document type	Journal Article
ZDB-ID	2827830-6
ISSN	2312-0541
ISSN	2312-0541
DOI	10.1183/23120541.00226-2018
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Article ; Online: Epithelial Yap/Taz are required for functional alveolar regeneration following acute lung injury.

DiGiovanni, Gianluca T / Han, Wei / Sherrill, Taylor P / Taylor, Chase J / Nichols, David S / Geis, Natalie M / Singha, Ujjal K / Calvi, Carla L / McCall, A Scott / Dixon, Molly M / Liu, Yang / Jang, Ji-Hoon / Gutor, Sergey S / Polosukhin, Vasiliy V / Blackwell, Timothy S / Kropski, Jonathan A / Gokey, Jason J

JCI insight

2023 Volume 8, Issue 19

Abstract: A hallmark of idiopathic pulmonary fibrosis (IPF) and other interstitial lung diseases is dysregulated repair of the alveolar epithelium. The Hippo pathway effector transcription factors YAP and TAZ are implicated as essential for type 1 and type 2 ... ...

Abstract	A hallmark of idiopathic pulmonary fibrosis (IPF) and other interstitial lung diseases is dysregulated repair of the alveolar epithelium. The Hippo pathway effector transcription factors YAP and TAZ are implicated as essential for type 1 and type 2 alveolar epithelial cell (AT1 and AT2) differentiation in the developing lung, yet aberrant activation of YAP/TAZ is a prominent feature of the dysregulated alveolar epithelium in IPF. In these studies, we sought to define the functional role of YAP/TAZ activity during alveolar regeneration. We demonstrated that Yap and Taz were normally activated in AT2 cells shortly after injury, and deletion of Yap/Taz in AT2 cells led to pathologic alveolar remodeling, failure of AT2-to-AT1 cell differentiation, increased collagen deposition, exaggerated neutrophilic inflammation, and increased mortality following injury induced by a single dose of bleomycin. Loss of Yap/Taz activity prior to an LPS injury prevented AT1 cell regeneration, led to intraalveolar collagen deposition, and resulted in persistent innate inflammation. These findings establish that AT2 cell Yap/Taz activity is essential for functional alveolar epithelial repair and prevention of fibrotic remodeling.
MeSH term(s)	Humans ; Acute Lung Injury ; Adaptor Proteins, Signal Transducing/genetics ; Adaptor Proteins, Signal Transducing/metabolism ; Collagen/metabolism ; Idiopathic Pulmonary Fibrosis/pathology ; Inflammation ; Regeneration ; Signal Transduction ; YAP-Signaling Proteins/metabolism
Chemical Substances	Adaptor Proteins, Signal Transducing ; Collagen (9007-34-5) ; YAP-Signaling Proteins ; WWTR1 protein, human
Language	English
Publishing date	2023-09-07
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.173374
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Article ; Online: Thromboxane-Prostanoid Receptor Signaling Drives Persistent Fibroblast Activation in Pulmonary Fibrosis.

American journal of respiratory and critical care medicine

2022 Volume 206, Issue 5, Page(s) 596–607

Abstract: Rationale: ...

Abstract	Rationale:
MeSH term(s)	Animals ; Bleomycin/pharmacology ; F2-Isoprostanes/metabolism ; Fibroblasts/metabolism ; Humans ; Idiopathic Pulmonary Fibrosis/genetics ; Lung/metabolism ; Mice ; Mice, Inbred C57BL ; Prostaglandins/metabolism ; Receptors, Thromboxane/metabolism ; Thromboxanes/metabolism ; Transforming Growth Factor beta/metabolism
Chemical Substances	F2-Isoprostanes ; Prostaglandins ; Receptors, Thromboxane ; Thromboxanes ; Transforming Growth Factor beta ; Bleomycin (11056-06-7)
Language	English
Publishing date	2022-06-18
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1180953-x
ISSN	1535-4970 ; 0003-0805 ; 1073-449X
ISSN (online)	1535-4970
ISSN	0003-0805 ; 1073-449X
DOI	10.1164/rccm.202106-1503OC
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Article ; Online: Gas6/MerTK signaling is negatively regulated by NF-κB and supports lung carcinogenesis.

Novitskiy, Sergey V / Zaynagetdinov, Rinat / Vasiukov, Georgii / Gutor, Sergey / Han, Wei / Serezani, Ana / Matafonov, Anton / Gleaves, Linda A / Sherrill, Taylor P / Polosukhin, Vasiliy V / Blackwell, Timothy S

Oncotarget

2019 Volume 10, Issue 66, Page(s) 7031–7042

Abstract: Growth arrest-specific 6 (Gas6) has been implicated in carcinogenesis through activation of its receptors, particularly MerTK. To investigate whether Gas6 plays a role in resistance to NF-κB inhibitors, which have not proven to be effective agents for ... ...

Abstract	Growth arrest-specific 6 (Gas6) has been implicated in carcinogenesis through activation of its receptors, particularly MerTK. To investigate whether Gas6 plays a role in resistance to NF-κB inhibitors, which have not proven to be effective agents for lung cancer therapy, we studied lung cancer models induced by urethane injection or expression of mutant Kras (Kras
Language	English
Publishing date	2019-12-17
Publishing country	United States
Document type	Journal Article
ZDB-ID	2560162-3
ISSN	1949-2553 ; 1949-2553
ISSN (online)	1949-2553
ISSN	1949-2553
DOI	10.18632/oncotarget.27345
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Article: Intraductal injection of lps as a mouse model of mastitis: signaling visualized via an nf-κb reporter transgenic

Barham, Whitney / Sherrill, Taylor / Connelly, Linda / Blackwell, Timothy S / Yull, Fiona E

Journal of visualized experiments. 2012 Sept. 04, , no. 67

2012

Abstract: Animal models of human disease are necessary in order to rigorously study stages of disease progression and associated mechanisms, and ultimately, as pre-clinical models to test interventions. In these methods, we describe a technique in which ... ...

Abstract	Animal models of human disease are necessary in order to rigorously study stages of disease progression and associated mechanisms, and ultimately, as pre-clinical models to test interventions. In these methods, we describe a technique in which lipopolysaccharide (LPS) is injected into the lactating mouse mammary gland via the nipple, effectively modeling mastitis, or inflammation, of the gland. This simulated infection results in increased nuclear factor kappa B (NF-κB) signaling, as visualized through bioluminescent imaging of an NF-κB luciferase reporter mouse1. Our ultimate goal in developing these methods was to study the inflammation associated with mastitis in the lactating gland, which often includes redness, swelling, and immune cell infiltration2,3. Therefore, we were keenly aware that incision or any type of wounding of the skin, the nipple, or the gland in order to introduce the LPS could not be utilized in our methods since the approach would likely confound the read-out of inflammation. We also desired a straight-forward method that did not require specially made hand-drawn pipettes or the use of micromanipulators to hold these specialized tools in place. Thus, we determined to use a commercially available insulin syringe and to inject the agent into the mammary duct of an intact nipple. This method was successful and allowed us to study the inflammation associated with LPS injection without any additional effects overlaid by the process of injection. In addition, this method also utilized an NF-κB luciferase reporter transgenic mouse and bioluminescent imaging technology to visually and quantitatively show increased NF-κB signaling within the LPS-injected gland4. These methods are of interest to researchers of many disciplines who wish to model disease within the lactating mammary gland, as ultimately, the technique described here could be utilized for injection of a number of substances, and is not limited to only LPS.
Keywords	animal models ; bioluminescence ; disease course ; human diseases ; image analysis ; inflammation ; insulin ; lactation ; lipopolysaccharides ; luciferase ; mammary glands ; mastitis ; mice ; transcription factor NF-kappa B ; transgenic animals
Language	English
Dates of publication	2012-0904
Size	p. e4030.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/4030
Database	NAL-Catalogue (AGRICOLA)

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