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Article ; Online: KCNF1 promotes lung cancer by modulating ITGB4 expression.

Chen, Ching-Yi / Wu, Pei-Ying / Van Scoyk, Michelle / Simko, Stephanie A / Chou, Chu-Fang / Winn, Robert A

2022 Volume 30, Issue 3, Page(s) 414–423

Abstract: Lung cancer continues to be the leading cause of cancer death in the United States. Despite recent advances, the five-year survival rate for lung cancer compared to other cancers still remains fairly low. The discovery of molecular targets for lung ... ...

Abstract	Lung cancer continues to be the leading cause of cancer death in the United States. Despite recent advances, the five-year survival rate for lung cancer compared to other cancers still remains fairly low. The discovery of molecular targets for lung cancer is key to the development of new approaches and therapies. Electrically silent voltage-gated potassium channel (KvS) subfamilies, which are unable to form functional homotetramers, are implicated in cell-cycle progression, cell proliferation and tumorigenesis. Here, we analyzed the expression of KvS subfamilies in human lung tumors and identified that potassium voltage-gated channel subfamily F member 1 (KCNF1) was up-regulated in non-small cell lung cancer (NSCLC). Silencing of KCNF1 in NSCLC cell lines reduced cell proliferation and tumor progression in mouse xenografts, re-established the integrity of the basement membrane, and enhanced cisplatin sensitivity. KCNF1 was predominately localized in the nucleoplasm and likely mediated its functions in an ion-independent manner. We identified integrin β4 subunit (ITGB4) as a downstream target for KCNF1. Our findings suggest that KCNF1 promotes lung cancer by enhancing ITGB4 signaling and implicate KCNF1 as a novel therapeutic target for lung cancer.
MeSH term(s)	Animals ; Humans ; Mice ; Carcinoma, Non-Small-Cell Lung/genetics ; Carcinoma, Non-Small-Cell Lung/pathology ; Cell Line, Tumor ; Cell Proliferation/genetics ; Gene Expression Regulation, Neoplastic ; Integrin beta4/genetics ; Integrin beta4/metabolism ; Lung Neoplasms/genetics ; Lung Neoplasms/pathology ; Signal Transduction
Chemical Substances	Integrin beta4 ; ITGB4 protein, human ; KCNF1 protein, human
Language	English
Publishing date	2022-11-16
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	1212513-1
ISSN	1476-5500 ; 0929-1903
ISSN (online)	1476-5500
ISSN	0929-1903
DOI	10.1038/s41417-022-00560-4
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Zs.A 4125: Show issues

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Article ; Online: Identification of the SARS-CoV-2 Entry Receptor ACE2 as a Direct Target for Transcriptional Repression by Miz1.

Yang, Jing / Perez, Edith A / Hou, Changchun / Zhang, Pin / Van Scoyk, Michelle / Winn, Robert A / Rong, Lijun / Liu, Jing

Frontiers in immunology

2021 Volume 12, Page(s) 648815

Abstract: Multiple lines of evidence have demonstrated that cigarette smoke or Chronic Obstructive Pulmonary Disease upregulates angiotensin-converting enzyme 2, the cellular receptor for the entry of the severe acute respiratory syndrome coronavirus 2, which ... ...

Abstract	Multiple lines of evidence have demonstrated that cigarette smoke or Chronic Obstructive Pulmonary Disease upregulates angiotensin-converting enzyme 2, the cellular receptor for the entry of the severe acute respiratory syndrome coronavirus 2, which predisposes individuals to develop severe Coronavirus disease 2019. The reason for this observation is unknown. We recently reported that the loss of function of Miz1 in the lung epithelium in mice leads to a spontaneous COPD-like phenotype, associated with upregulation of angiotensin-converting enzyme 2. We also reported that cigarette smoke exposure downregulates Miz1 in lung epithelial cells and in mice, and Miz1 is also downregulated in the lungs of COPD patients. Here, we provide further evidence that Miz1 directly binds to and represses the promoter of angiotensin-converting enzyme 2 in mouse and human lung epithelial cells. Our data provide a potential molecular mechanism for the upregulation of angiotensin-converting enzyme 2 observed in smokers and COPD patients, with implication in severe Coronavirus disease 2019.
MeSH term(s)	Alveolar Epithelial Cells/metabolism ; Angiotensin-Converting Enzyme 2/genetics ; Angiotensin-Converting Enzyme 2/metabolism ; Animals ; BTB-POZ Domain ; Cell Line ; Cigarette Smoking/adverse effects ; Kruppel-Like Transcription Factors/chemistry ; Kruppel-Like Transcription Factors/genetics ; Kruppel-Like Transcription Factors/metabolism ; Mice ; Promoter Regions, Genetic ; Protein Binding ; Receptors, Virus/genetics ; Receptors, Virus/metabolism ; SARS-CoV-2/physiology ; Spike Glycoprotein, Coronavirus/metabolism ; Transcription, Genetic/drug effects ; Tumor Necrosis Factors/pharmacology ; Virus Internalization
Chemical Substances	Kruppel-Like Transcription Factors ; Receptors, Virus ; Spike Glycoprotein, Coronavirus ; Tumor Necrosis Factors ; ZBTB17 protein, human ; spike protein, SARS-CoV-2 ; ACE2 protein, human (EC 3.4.17.23) ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23)
Language	English
Publishing date	2021-07-07
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2021.648815
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Article ; Online: Cooperation between PRMT1 and PRMT6 drives lung cancer health disparities among Black/African American men.

iScience

2024 Volume 27, Issue 2, Page(s) 108858

Abstract: Lung cancer is the third most common cancer with Black/AA men showing higher risk and poorer outcomes than NHW men. Lung cancer disparities are multifactorial, driven by tobacco exposure, inequities in care access, upstream health determinants, and ... ...

Abstract	Lung cancer is the third most common cancer with Black/AA men showing higher risk and poorer outcomes than NHW men. Lung cancer disparities are multifactorial, driven by tobacco exposure, inequities in care access, upstream health determinants, and molecular determinants including biological and genetic factors. Elevated expressions of protein arginine methyltransferases (PRMTs) correlating with poorer prognosis have been observed in many cancers. Most importantly, our study shows that PRMT6 displays higher expression in lung cancer tissues of Black/AA men compared to NHW men. In this study, we investigated the underlying mechanism of PRMT6 and its cooperation with PRMT1 to form a heteromer as a driver of lung cancer. Disrupting PRMT1/PRMT6 heteromer by a competitive peptide reduced proliferation in non-small cell lung cancer cell lines and patient-derived organoids, therefore, giving rise to a more strategic approach in the treatment of Black/AA men with lung cancer and to eliminate cancer health disparities.
Language	English
Publishing date	2024-01-11
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2024.108858
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Article ; Online: Miz1 promotes KRAS-driven lung tumorigenesis by repressing the protocadherin Pcdh10.

Yang, Jing / Hou, Changchun / Wang, Huashan / Perez, Edith A / Do-Umehara, Hanh Chi / Dong, Huali / Arunagiri, Vinothini / Tong, Fangjia / Van Scoyk, Michelle / Cho, Minsu / Liu, Xinyi / Ge, Xiaodong / Winn, Robert A / Ridge, Karen M / Wang, Xiaowei / Chandel, Navdeep S / Liu, Jing

Cancer letters

2022 Volume 555, Page(s) 216025

Abstract: Targeting KRAS-mutated non-small-cell lung cancer (NSCLC) remains clinically challenging. Here we show that loss of function of Miz1 inhibits lung tumorigenesis in a mouse model of oncogenic KRAS-driven lung cancer. In vitro, knockout or silencing of ... ...

Abstract	Targeting KRAS-mutated non-small-cell lung cancer (NSCLC) remains clinically challenging. Here we show that loss of function of Miz1 inhibits lung tumorigenesis in a mouse model of oncogenic KRAS-driven lung cancer. In vitro, knockout or silencing of Miz1 decreases cell proliferation, clonogenicity, migration, invasion, or anchorage-independent growth in mutant (MT) KRAS murine or human NSCLC cells but has unremarkable impact on non-tumorigenic cells or wild-type (WT) KRAS human NSCLC cells. RNA-sequencing reveals Protocadherin-10 (Pcdh10) as the top upregulated gene by Miz1 knockout in MT KRAS murine lung tumor cells. Chromatin immunoprecipitation shows Miz1 binding on the Pcdh10 promoter in MT KRAS lung tumor cells but not non-tumorigenic cells. Importantly, silencing of Pcdh10 rescues cell proliferation and clonogenicity in Miz1 knockout/knockdown MT KRAS murine or human tumor cells, and rescues allograft tumor growth of Miz1 knockout tumor cells in vivo. Miz1 is upregulated in MT KRAS lung tumor tissues compared with adjacent non-involved tissues in mice. Consistent with this, Miz1 is upregulated while Pcdh10 is downregulated in human lung adenocarcinomas (LUAD) compared with normal tissues, and high Miz1 levels or low Pcdh10 levels are associated with poor survival in lung cancer patients. Furthermore, the Miz1 signature is associated with worse survival in MT but not WT KRAS LUAD, and Pcdh10 is downregulated in MT compared to WT KRAS LUAD. Taken together, our studies implicate the Miz1/Pcdh10 axis in oncogenic KRAS-driven lung tumorigenesis.
MeSH term(s)	Animals ; Humans ; Mice ; Carcinogenesis/genetics ; Carcinoma, Non-Small-Cell Lung/pathology ; Cell Line, Tumor ; Lung/pathology ; Lung Neoplasms/metabolism ; Protein Inhibitors of Activated STAT/metabolism ; Proto-Oncogene Proteins p21(ras)/genetics ; Proto-Oncogene Proteins p21(ras)/metabolism ; Protocadherins ; Ubiquitin-Protein Ligases/metabolism
Chemical Substances	KRAS protein, human ; Miz1 protein, mouse (EC 2.3.2.27) ; PCDH10 protein, human ; Pcdh10 protein, mouse ; Protein Inhibitors of Activated STAT ; Proto-Oncogene Proteins p21(ras) (EC 3.6.5.2) ; Protocadherins ; Ubiquitin-Protein Ligases (EC 2.3.2.27) ; Hras protein, mouse (EC 3.6.5.2) ; ZBTB17 protein, human
Language	English
Publishing date	2022-12-17
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	195674-7
ISSN	1872-7980 ; 0304-3835
ISSN (online)	1872-7980
ISSN	0304-3835
DOI	10.1016/j.canlet.2022.216025
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Zs.A 1177: Show issues

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Article ; Online: PRMT6 Promotes Lung Tumor Progression via the Alternate Activation of Tumor-Associated Macrophages.

Avasarala, Sreedevi / Wu, Pei-Ying / Khan, Samia Q / Yanlin, Su / Van Scoyk, Michelle / Bao, Jianqiang / Di Lorenzo, Alessandra / David, Odile / Bedford, Mark T / Gupta, Vineet / Winn, Robert A / Bikkavilli, Rama Kamesh

Molecular cancer research : MCR

2019 Volume 18, Issue 1, Page(s) 166–178

Abstract: Increased expression of protein arginine methyl transferase 6 (PRMT6) correlates with worse prognosis in lung cancer cases. To interrogate ... ...

Abstract	Increased expression of protein arginine methyl transferase 6 (PRMT6) correlates with worse prognosis in lung cancer cases. To interrogate the
MeSH term(s)	Animals ; Cell Culture Techniques ; Cell Line, Tumor ; Cell Proliferation ; Disease Models, Animal ; Disease Progression ; Humans ; Lung Neoplasms/genetics ; Lung Neoplasms/mortality ; Lung Neoplasms/pathology ; Macrophages/metabolism ; Mice ; Nuclear Proteins/genetics ; Protein-Arginine N-Methyltransferases/genetics ; Survival Analysis
Chemical Substances	Nuclear Proteins ; PRMT6 protein, human (EC 2.1.1.319) ; Protein-Arginine N-Methyltransferases (EC 2.1.1.319)
Language	English
Publishing date	2019-10-16
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	2098788-2
ISSN	1557-3125 ; 1541-7786
ISSN (online)	1557-3125
ISSN	1541-7786
DOI	10.1158/1541-7786.MCR-19-0204
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Zs.A 5744: Show issues

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Article ; Online: Role of the wnt signaling pathway and lung cancer.

Tennis, Meredith / Van Scoyk, Michelle / Winn, Robert A

Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer

2007 Volume 2, Issue 10, Page(s) 889–892

MeSH term(s)	Animals ; Humans ; Lung Neoplasms/etiology ; Lung Neoplasms/metabolism ; Lung Neoplasms/therapy ; Signal Transduction ; Wnt Proteins/metabolism
Chemical Substances	Wnt Proteins
Language	English
Publishing date	2007-10
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	2432037-7
ISSN	1556-1380 ; 1556-0864
ISSN (online)	1556-1380
ISSN	1556-0864
DOI	10.1097/JTO.0b013e318153fdb1
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Zs.A 6664: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular ab Jg. 2022: Lesesaal (EG)
Zs.MO 652: Show issues

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Article ; Online: Lysocardiolipin acyltransferase regulates NSCLC cell proliferation and migration by modulating mitochondrial dynamics.

Huang, Long Shuang / Kotha, Sainath R / Avasarala, Sreedevi / VanScoyk, Michelle / Winn, Robert A / Pennathur, Arjun / Yashaswini, Puttaraju S / Bandela, Mounica / Salgia, Ravi / Tyurina, Yulia Y / Kagan, Valerian E / Zhu, Xiangdong / Reddy, Sekhar P / Sudhadevi, Tara / Punathil-Kannan, Prasanth-Kumar / Harijith, Anantha / Ramchandran, Ramaswamy / Bikkavilli, Rama Kamesh / Natarajan, Viswanathan

The Journal of biological chemistry

2020 Volume 295, Issue 38, Page(s) 13393–13406

Abstract: Lysocardiolipin acyltransferase (LYCAT), a cardiolipin (CL)-remodeling enzyme, is crucial for maintaining normal mitochondrial function and vascular development. Despite the well-characterized role for LYCAT in the regulation of mitochondrial dynamics, ... ...

Abstract	Lysocardiolipin acyltransferase (LYCAT), a cardiolipin (CL)-remodeling enzyme, is crucial for maintaining normal mitochondrial function and vascular development. Despite the well-characterized role for LYCAT in the regulation of mitochondrial dynamics, its involvement in lung cancer, if any, remains incompletely understood. In this study,
MeSH term(s)	1-Acylglycerol-3-Phosphate O-Acyltransferase/genetics ; 1-Acylglycerol-3-Phosphate O-Acyltransferase/metabolism ; A549 Cells ; Animals ; Carcinoma, Non-Small-Cell Lung/enzymology ; Carcinoma, Non-Small-Cell Lung/genetics ; Carcinoma, Non-Small-Cell Lung/pathology ; Cardiolipins/genetics ; Cardiolipins/metabolism ; Cell Proliferation ; Heterografts ; Humans ; Lung Neoplasms/enzymology ; Lung Neoplasms/genetics ; Lung Neoplasms/pathology ; Mice ; Mice, Nude ; Mitochondria/genetics ; Mitochondria/metabolism ; Neoplasm Proteins/genetics ; Neoplasm Proteins/metabolism ; Neoplasm Transplantation
Chemical Substances	Cardiolipins ; Neoplasm Proteins ; 1-Acylglycerol-3-Phosphate O-Acyltransferase (EC 2.3.1.51) ; LCLAT1 protein, human (EC 2.3.1.51)
Language	English
Publishing date	2020-07-30
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.RA120.012680
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Uc I Zs.108: Show issues

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Article ; Online: In vitro methylation assay to study protein arginine methylation.

Bikkavilli, Rama Kamesh / Avasarala, Sreedevi / Van Scoyk, Michelle / Karuppusamy Rathinam, Manoj Kumar / Tauler, Jordi / Borowicz, Stanley / Winn, Robert A

Journal of visualized experiments : JoVE

2014 , Issue 92, Page(s) e51997

Abstract: Protein arginine methylation is one of the most abundant post-translational modifications in the nucleus. Protein arginine methylation can be identified and/or determined via proteomic approaches, and/or immunoblotting with methyl-arginine specific ... ...

Abstract	Protein arginine methylation is one of the most abundant post-translational modifications in the nucleus. Protein arginine methylation can be identified and/or determined via proteomic approaches, and/or immunoblotting with methyl-arginine specific antibodies. However, these techniques sometimes can be misleading and often provide false positive results. Most importantly, these techniques cannot provide direct evidence in support of the PRMT substrate specificity. In vitro methylation assays, on the other hand, are useful biochemical assays, which are sensitive, and consistently reveal if the identified proteins are indeed PRMT substrates. A typical in vitro methylation assay includes purified, active PRMTs, purified substrate and a radioisotope labeled methyl donor (S-adenosyl-L-[methyl-(3)H] methionine). Here we describe a step-by-step protocol to isolate catalytically active PRMT1, a ubiquitously expressed PRMT family member. The methyl transferase activities of the purified PRMT1 were later tested on Ras-GTPase activating protein binding protein 1 (G3BP1), a known PRMT substrate, in the presence of S-adenosyl-L-[methyl-(3)H] methionine as the methyl donor. This protocol can be employed not only for establishing the methylation status of novel physiological PRMT1 substrates, but also for understanding the basic mechanism of protein arginine methylation.
MeSH term(s)	Arginine/chemistry ; Arginine/metabolism ; Carrier Proteins/chemistry ; Carrier Proteins/metabolism ; DNA Helicases ; Humans ; Methylation ; Poly-ADP-Ribose Binding Proteins ; Protein-Arginine N-Methyltransferases/chemistry ; Protein-Arginine N-Methyltransferases/metabolism ; RNA Helicases ; RNA Recognition Motif Proteins ; S-Adenosylmethionine/analogs & derivatives ; S-Adenosylmethionine/chemistry ; S-Adenosylmethionine/metabolism
Chemical Substances	Carrier Proteins ; Poly-ADP-Ribose Binding Proteins ; RNA Recognition Motif Proteins ; S-adenosyl-2-methylmethionine (16720-69-7) ; S-Adenosylmethionine (7LP2MPO46S) ; Arginine (94ZLA3W45F) ; Protein-Arginine N-Methyltransferases (EC 2.1.1.319) ; DNA Helicases (EC 3.6.4.-) ; G3BP1 protein, human (EC 3.6.4.12) ; RNA Helicases (EC 3.6.4.13)
Language	English
Publishing date	2014-10-05
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/51997
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Article ; Online: The soft agar colony formation assay.

Borowicz, Stanley / Van Scoyk, Michelle / Avasarala, Sreedevi / Karuppusamy Rathinam, Manoj Kumar / Tauler, Jordi / Bikkavilli, Rama Kamesh / Winn, Robert A

Journal of visualized experiments : JoVE

2014 , Issue 92, Page(s) e51998

Abstract: Anchorage-independent growth is the ability of transformed cells to grow independently of a solid surface, and is a hallmark of carcinogenesis. The soft agar colony formation assay is a well-established method for characterizing this capability in vitro ... ...

Abstract	Anchorage-independent growth is the ability of transformed cells to grow independently of a solid surface, and is a hallmark of carcinogenesis. The soft agar colony formation assay is a well-established method for characterizing this capability in vitro and is considered to be one of the most stringent tests for malignant transformation in cells. This assay also allows for semi-quantitative evaluation of this capability in response to various treatment conditions. Here, we will demonstrate the soft agar colony formation assay using a murine lung carcinoma cell line, CMT167, to demonstrate the tumor suppressive effects of two members of the Wnt signaling pathway, Wnt7A and Frizzled-9 (Fzd-9). Concurrent overexpression of Wnt7a and Fzd-9 caused an inhibition of colony formation in CMT167 cells. This shows that expression of Wnt7a ligand and its Frizzled-9 receptor is sufficient to suppress tumor growth in a murine lung carcinoma model.
MeSH term(s)	Animals ; Cell Line, Tumor ; Colony-Forming Units Assay/methods ; Lung Neoplasms/pathology ; Mice ; Tumor Stem Cell Assay/methods
Language	English
Publishing date	2014-10-27
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Video-Audio Media
ISSN	1940-087X
ISSN (online)	1940-087X
DOI	10.3791/51998
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: In vitro methylation assay to study protein arginine methylation

Bikkavilli, Rama Kamesh / Avasarala, Sreedevi / Van Scoyk, Michelle / Karuppusamy Rathinam, Manoj Kumar / Tauler, Jordi / Borowicz, Stanley / Winn, Robert A

Journal of visualized experiments. 2014 Oct. 05, , no. 92

2014

Abstract	Protein arginine methylation is one of the most abundant post-translational modifications in the nucleus. Protein arginine methylation can be identified and/or determined via proteomic approaches, and/or immunoblotting with methyl-arginine specific antibodies. However, these techniques sometimes can be misleading and often provide false positive results. Most importantly, these techniques cannot provide direct evidence in support of the PRMT substrate specificity. In vitro methylation assays, on the other hand, are useful biochemical assays, which are sensitive, and consistently reveal if the identified proteins are indeed PRMT substrates. A typical in vitro methylation assay includes purified, active PRMTs, purified substrate and a radioisotope labeled methyl donor (S-adenosyl-L-[methyl-3H] methionine). Here we describe a step-by-step protocol to isolate catalytically active PRMT1, a ubiquitously expressed PRMT family member. The methyl transferase activities of the purified PRMT1 were later tested on Ras-GTPase activating protein binding protein 1 (G3BP1), a known PRMT substrate, in the presence of S-adenosyl-L-[methyl-3H] methionine as the methyl donor. This protocol can be employed not only for establishing the methylation status of novel physiological PRMT1 substrates, but also for understanding the basic mechanism of protein arginine methylation.
Keywords	antibodies ; arginine ; binding proteins ; enzyme activity ; false positive results ; immunoblotting ; methionine ; methylation ; post-translational modification ; proteomics ; substrate specificity ; type I protein arginine methyltransferase
Language	English
Dates of publication	2014-1005
Size	p. e51997.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/51997
Database	NAL-Catalogue (AGRICOLA)

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