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Article ; Online: Unilateral Lung Removal in Combination with Monocrotaline or SU5416 in Rodents: A Reliable Model to Mimic the Pathology of the Human Pulmonary Hypertension.

Katz, Michael G / Hadas, Yoav / Shtraizent, Nataly / Ravvin, Shana / Madjarov, Jeko M / Eliyahu, Efrat

Methods in molecular biology (Clifton, N.J.)

2024 Volume 2803, Page(s) 173–185

Abstract: Pulmonary hypertension (PH) is a chronic and progressive disorder characterized by elevated mean pulmonary arterial pressure, pulmonary vascular remodeling, and the development of concentric laminar intimal fibrosis with plexiform lesions. While rodent ... ...

Abstract	Pulmonary hypertension (PH) is a chronic and progressive disorder characterized by elevated mean pulmonary arterial pressure, pulmonary vascular remodeling, and the development of concentric laminar intimal fibrosis with plexiform lesions. While rodent models have been developed to study PH, they have certain deficiencies and do not entirely replicate the human disease due to the heterogeneity of PH pathology. Therefore, combined models are necessary to study PH. Recent studies have shown that altered pulmonary blood flow is a significant trigger in the development of vascular remodeling and neointimal lesions. One of the most promising rodent models for increased pulmonary flow is the combination of unilateral left pneumonectomy with a "second hit" of monocrotaline (MCT) or SU5416. The removal of one lung in this model forces blood to circulate only in the other lung and induces increased and turbulent pulmonary blood flow. This increased vascular flow leads to progressive remodeling and occlusion of small pulmonary arteries. The second hit by MCT or SU5416 leads to endothelial cell dysfunction, resulting in severe PH and the development of plexiform arteriopathy.
MeSH term(s)	Hypertension, Pulmonary/pathology ; Hypertension, Pulmonary/etiology ; Hypertension, Pulmonary/chemically induced ; Monocrotaline ; Indoles ; Disease Models, Animal ; Animals ; Rats ; Humans ; Pyrroles ; Lung/pathology ; Pneumonectomy/methods ; Vascular Remodeling ; Pulmonary Artery/pathology ; Mice
Chemical Substances	Monocrotaline (73077K8HYV) ; Indoles ; Semaxinib (71IA9S35AJ) ; Pyrroles
Language	English
Publishing date	2024-04-27
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3846-0_13
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Cardiac Targeted Adeno-Associated Virus Injection in Rats.

Katz, Michael G / Hadas, Yoav / Vincek, Adam S / Shtraizent, Nataly / Schadt, Eric / Eliyahu, Efrat

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2573, Page(s) 135–145

Abstract: Gene therapy is a promising approach in the treatment of cardiovascular diseases. The vectors available for cardiovascular gene therapy have significantly improved over time. Cardiac tropism is a primary characteristic of an ideal vector along with a ... ...

Abstract	Gene therapy is a promising approach in the treatment of cardiovascular diseases. The vectors available for cardiovascular gene therapy have significantly improved over time. Cardiac tropism is a primary characteristic of an ideal vector along with a long-term expression profile and a minimal risk of cellular immune response. Preclinical and clinical studies have demonstrated that adeno-associated viral (AAV) vectors are one of the most attractive vehicles for gene transfer. AAV has gained great popularity in the last years because of its biological properties and advantages over other viral vector systems. In this chapter we will describe methods for intracardiac delivery of AAV vector in rats.
MeSH term(s)	Animals ; Dependovirus/genetics ; Gene Transfer Techniques ; Genetic Therapy/methods ; Genetic Vectors/genetics ; Heart ; Rats
Language	English
Publishing date	2022-08-30
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2707-5_10
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Article ; Online: Acid ceramidase gene therapy ameliorates pulmonary arterial hypertension with right heart dysfunction.

Katz, Michael G / Hadas, Yoav / Vincek, Adam / Freage-Kahn, Lina / Shtraizent, Nataly / Madjarov, Jeko M / Pastuszko, Peter / Eliyahu, Efrat

Respiratory research

2023 Volume 24, Issue 1, Page(s) 197

Abstract: Background: Up-regulation of ceramides in pulmonary hypertension (PH), contributing to perturbations in sphingolipid homeostasis and the transition of cells to a senescence state. We assessed the safety, feasibility, and efficiency of acid ceramidase ... ...

Abstract	Background: Up-regulation of ceramides in pulmonary hypertension (PH), contributing to perturbations in sphingolipid homeostasis and the transition of cells to a senescence state. We assessed the safety, feasibility, and efficiency of acid ceramidase gene transfer in a rodent PH model. Methods: A model of PH was established by the combination of left pneumonectomy and injection of Sugen toxin. Magnetic resonance imaging and right heart catheterization confirmed development of PH. Animals were subjected to intratracheal administration of synthetic adeno-associated viral vector (Anc80L65) carrying the acid ceramidase (Anc80L65.AC), an empty capsid vector, or saline. Therapeutic efficacy was evaluated 8 weeks after gene delivery. Results: Hemodynamic assessment 4 weeks after PH model the development demonstrated an increase in the mean pulmonary artery pressure to 30.4 ± 2.13 mmHg versus 10.4 ± 1.65 mmHg in sham (p < 0.001), which was consistent with the definition of PH. We documented a significant increase in pulmonary vascular resistance in the saline-treated (6.79 ± 0.85 mm Hg) and empty capsid (6.94 ± 0.47 mm Hg) groups, but not in animals receiving Anc80L65.AC (4.44 ± 0.71 mm Hg, p < 0.001). Morphometric analysis demonstrated an increase in medial wall thickness in control groups in comparison to those treated with acid ceramidase. After acid ceramidase gene delivery, a significant decrease of pro-inflammatory factors, interleukins, and senescence markers was observed. Conclusion: Gene delivery of acid ceramidase provided tropism to pulmonary tissue and ameliorated vascular remodeling with right ventricular dysfunction in pulmonary hypertension.
MeSH term(s)	Animals ; Pulmonary Arterial Hypertension ; Hypertension, Pulmonary/genetics ; Hypertension, Pulmonary/therapy ; Hypertension, Pulmonary/pathology ; Acid Ceramidase/genetics ; Familial Primary Pulmonary Hypertension ; Genetic Therapy ; Pulmonary Artery/pathology ; Ventricular Dysfunction, Right
Chemical Substances	Acid Ceramidase (EC 3.5.1.23)
Language	English
Publishing date	2023-08-11
Publishing country	England
Document type	Journal Article
ZDB-ID	2041675-1
ISSN	1465-993X ; 1465-993X
ISSN (online)	1465-993X
ISSN	1465-993X
DOI	10.1186/s12931-023-02487-2
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Article ; Online: Surgical Methods for Cardiac Gene Delivery in Large Animals.

Katz, Michael G / Hadas, Yoav / Vincek, Adam S / Shtraizent, Nataly / Gordon, Hylton P / Pastuszko, Peter / Schadt, Eric / Eliyahu, Efrat

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2573, Page(s) 189–203

Abstract: This chapter describes main strategies of surgical gene delivery in large animals. Existing methods of cardiac gene transfer can be classified by the site of injection, interventional approach, and type of cardiac circulation at the time of transfer. ... ...

Abstract	This chapter describes main strategies of surgical gene delivery in large animals. Existing methods of cardiac gene transfer can be classified by the site of injection, interventional approach, and type of cardiac circulation at the time of transfer. Randomized clinical trials have suggested that the therapeutic benefits of gene therapy are not as substantial as expected from animal studies. This discordance in results is largely due to gene delivery methods that may be effective in small animals but are not scalable to larger species and, therefore, cannot transduce a sufficient fraction of myocytes to establish long-term clinical efficacy. Ideally, an optimized gene transfer should incorporate the following: a closed-loop recirculation for extended transgene residence time; vector washout form the vascular system after transfer to prevent collateral expression; use of methods to increase myocardial transcapillary gradient for viral particles for a better transduction, probably retrograde route of gene delivery through the coronary venous system; and myocardial ischemic preconditioning.
MeSH term(s)	Animals ; Gene Transfer Techniques ; Genetic Therapy/methods ; Genetic Vectors/genetics ; Injections ; Myocardium/metabolism ; Transgenes
Language	English
Publishing date	2022-08-30
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2707-5_15
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Article ; Online: Effects of Hormone Replacement Therapy on Women's Lung Health and Disease.

Eliyahu, Efrat / Katz, Michael G / Vincek, Adam / Freage-Kahn, Lina / Ravvin, Shana / Tal, Smadar / Grage, Henry / Shtraizent, Nataly / Barak, Tuvia / Arkush, Bezalel

Pulmonary therapy

2023 Volume 9, Issue 4, Page(s) 461–477

Abstract: This review provides an overview of menopausal hormone therapy and pulmonary disease risk, with a focus on the effect of hormone replacement therapy (HRT) on pulmonary function and its relation to lung diseases. This summary is based on authors' ... ...

Abstract	This review provides an overview of menopausal hormone therapy and pulmonary disease risk, with a focus on the effect of hormone replacement therapy (HRT) on pulmonary function and its relation to lung diseases. This summary is based on authors' knowledge in the field of HRT and supplemented by a PubMed search using the terms "menopause hormone therapy," "asthma", "lung cancer", "chronic obstructive pulmonary disease", "lung function", and "pulmonary hypertension". Available evidence indicates that there is limited research on the role of sex hormones in the susceptibility, severity, and progression of chronic respiratory diseases. However, some studies suggest that the hormonal changes that occur during the menopausal transition may have an impact on pulmonary function and respiratory diseases. Women are in need of convenient access to a safe and effective modality for personalized HRT based on an artificial intelligence (AI)-driven platform that will enable them to receive personalized hormonal treatment through frequent, convenient, and accurate measurements of hormone levels in peripheral blood.
Language	English
Publishing date	2023-10-10
Publishing country	United States
Document type	Journal Article ; Review
ISSN	2364-1746
ISSN (online)	2364-1746
DOI	10.1007/s41030-023-00240-0
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Article ; Online: Chronic thoracic pain after cardiac surgery: role of inflammation and biomechanical sternal stability.

Madjarov, Jeko M / Katz, Michael G / Hadas, Yoav / Kim, Sofia Jisoo / Freage-Kahn, Lina / Madzharov, Svetozar / Vincek, Adam / Madjarova, Sophia J / Seidman, Piers / Shtraizent, Nataly / Robicsek, Steven A / Eliyahu, Efrat

Frontiers in pain research (Lausanne, Switzerland)

2023 Volume 4, Page(s) 1180969

Abstract: Introduction: The pathogenesis of chronic chest pain after cardiac surgery has not been determinate. If left untreated, postoperative sternal pain reduces the quality of life and patient satisfaction with cardiac surgery. The purpose of the study was to ...

Abstract	Introduction: The pathogenesis of chronic chest pain after cardiac surgery has not been determinate. If left untreated, postoperative sternal pain reduces the quality of life and patient satisfaction with cardiac surgery. The purpose of the study was to examine the effect of chest inflammation on postoperative pain, risk factors for chronic pain after cardiac surgery and to explore how chest reconstruction was associated with the intensity of pain. Methods: The authors performed a study of acute and chronic thoracic pain after cardiac surgery in patients with and without sternal infection and compared different techniques for chest reconstruction. 42 high-risk patients for the development of mediastinitis were included. Patients with mediastinitis received chest reconstruction (group 1). Their demographics and risk factors were matched with no-infection patients with chest reconstruction (group 2) and subjects who underwent conventional sternal closure (group 3). Chronic pain was assessed by the numeric rating scale after surgery. Results: The assessment of the incidence and intensity of chest pain at 3 months post-surgery demonstrated that 14 out of 42 patients across all groups still experienced chronic pain. Specifically, in group 1 with sternal infection five patients had mild pain, while one patient experienced mild pain in group 2, and eight patients in group 3. Also, follow-up results indicated that the highest pain score was in group 3. While baseline levels of cytokines were increased among patients with sternal infection, at discharge only the level of interleukin 6 remained high compared to no infection groups. Compared to conventional closure, after chest reconstruction, we found better healing scores at 3-month follow-up and a higher percentage of patients with the complete sternal union. Conclusions: Overall, 14 out of 42 patients have chronic pain after cardiac surgery. The intensity of the pain in mediastinitis patients significantly decreased at 3 months follow-up after chest reconstruction. Thus, post-surgery mediastinitis is not a determining factor for development the chronic chest pain. There is no correlation between cytokines levels and pain score except interleukin 6 which remains elevated for a long time after treatment. Correlation between sternal healing score and chronic chest pain was demonstrated.
Language	English
Publishing date	2023-08-11
Publishing country	Switzerland
Document type	Journal Article
ISSN	2673-561X
ISSN (online)	2673-561X
DOI	10.3389/fpain.2023.1180969
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Article ; Online: Efficient cardiac gene transfer and early-onset expression of a synthetic adeno-associated viral vector, Anc80L65, after intramyocardial administration.

Katz, Michael G / Hadas, Yoav / Bailey, Rasheed A / Fazal, Shahood / Vincek, Adam / Madjarova, Sophia J / Shtraizent, Nataly / Vandenberghe, Luk H / Eliyahu, Efrat

The Journal of thoracic and cardiovascular surgery

2021 Volume 164, Issue 6, Page(s) e429–e443

Abstract: Objective: Gene therapy is a promising approach in the treatment of cardiovascular diseases. Preclinical and clinical studies have demonstrated that adeno-associated viral vectors are the most attractive vehicles for gene transfer. However, preexisting ... ...

Abstract	Objective: Gene therapy is a promising approach in the treatment of cardiovascular diseases. Preclinical and clinical studies have demonstrated that adeno-associated viral vectors are the most attractive vehicles for gene transfer. However, preexisting immunity, delayed gene expression, and postinfection immune response limit the success of this technology. The aim of this study was to investigate the efficacy of the first synthetic adeno-associated viral lineage clone, Anc80L65, for cardiac gene therapy. Methods: By combining 2 different reporter approaches by fluorescence with green fluorescent protein and bioluminescence (Firefly luciferase), we compared transduction efficiency of Anc80L65 and adeno-associated virus, serotype 9 in neonatal rat cardiomyocytes ex vivo and rat hearts in vivo after intramyocardial and intracoronary administration. Results: In cardiomyocytes, Anc80L65 provided a green fluorescent protein expression of 28.9% (36.4 ± 3.34 cells/field) at 24 hours and approximately 100% on day 7. In contrast, adeno-associated virus, serotype 9 green fluorescent protein provided minimal green fluorescent protein expression of 5.64% at 24 hours and 11.8% on day 7. After intramyocardial injection, vector expression peaked on day 7 with Anc80L65; however, with adeno-associated virus, serotype 9 the peak expression was during week 6. Administration of Anc80L65 demonstrated significantly more efficient expression of reporter gene than after adeno-associated virus, serotype 9 at 6 weeks (6.81 ± 0.64 log Conclusions: Anc80L65 vector allows fast and robust gene transduction compared with adeno-associated virus, serotype 9 vector in cardiac gene therapy. Anc80L65 did not adversely affect cardiac function and caused no inflammatory response or toxicity.
MeSH term(s)	Rats ; Animals ; Genetic Vectors ; Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism ; Dependovirus/genetics ; Genetic Therapy/methods ; Myocytes, Cardiac/metabolism ; Gene Transfer Techniques ; Transduction, Genetic
Chemical Substances	Green Fluorescent Proteins (147336-22-9)
Language	English
Publishing date	2021-06-17
Publishing country	United States
Document type	Journal Article
ZDB-ID	3104-5
ISSN	1097-685X ; 0022-5223
ISSN (online)	1097-685X
ISSN	0022-5223
DOI	10.1016/j.jtcvs.2021.05.050
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Article ; Online: Hot Spot Mutation in TP53 (R248Q) Causes Oncogenic Gain-of-Function Phenotypes in a Breast Cancer Cell Line Derived from an African American patient.

Shtraizent, Nataly / Matsui, Hiroshi / Polotskaia, Alla / Bargonetti, Jill

International journal of environmental research and public health

2015 Volume 13, Issue 1, Page(s) ijerph13010022

Abstract: African American (AA) breast cancer patients often have triple negative breast cancer (TNBC) that contains mutations in the TP53 gene. The point mutations at amino acid residues R273 and R248 both result in oncogenic gain-of-function (GOF) phenotypes. ... ...

Abstract	African American (AA) breast cancer patients often have triple negative breast cancer (TNBC) that contains mutations in the TP53 gene. The point mutations at amino acid residues R273 and R248 both result in oncogenic gain-of-function (GOF) phenotypes. Expression of mutant p53 (mtp53) R273H associates with increased cell elasticity, survival under serum deprivation conditions, and increased Poly (ADP ribose) polymerase 1 (PARP1) on the chromatin in the AA-derived TNBC breast cancer cell line MDA-MB-468. We hypothesized that GOF mtp53 R248Q expression could stimulate a similar phenotype in the AA-derived TNBC cell line HCC70. To test this hypothesis we depleted the R248Q protein in the HCC70 cell line using shRNA-mediated knockdown. Using impedance-based real-time analysis we correlated the expression of mtp53 R248Q with increased cell deformability. We also documented that depletion of mtp53 R248Q increased PARP1 in the cytoplasm and decreased PARP1 on the chromatin. We conclude that in the AA-derived TNBC HCC70 cells mtp53 R248Q expression results in a causative tumor associated phenotype. This study supports using the biological markers of high expression of mtp53 R273H or R248Q as additional diagnostics for TNBC resistant subtypes often found in the AA community. Each mtp53 protein must be considered separately and this work adds R248Q to the increasing list of p53 mutations that can be used for diagnostics and drug targeting. Here we report that when R248Q mtp53 proteins are expressed in TNBC, then targeting the gain-of-function pathways may improve treatment efficacy.
MeSH term(s)	Adult ; African Americans/genetics ; Aged ; Aged, 80 and over ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Middle Aged ; Mutation/genetics ; Phenotype ; Triple Negative Breast Neoplasms/genetics ; Tumor Suppressor Protein p53/genetics
Chemical Substances	Tumor Suppressor Protein p53
Language	English
Publishing date	2015-12-22
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1660-4601
ISSN (online)	1660-4601
DOI	10.3390/ijerph13010022
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Article ; Online: Hot Spot Mutation in TP53 (R248Q) Causes Oncogenic Gain-of-Function Phenotypes in a Breast Cancer Cell Line Derived from an African American patient

Nataly Shtraizent / Hiroshi Matsui / Alla Polotskaia / Jill Bargonetti

International Journal of Environmental Research and Public Health, Vol 13, Iss 1, p

2015 Volume 22

Abstract	African American (AA) breast cancer patients often have triple negative breast cancer (TNBC) that contains mutations in the TP53 gene. The point mutations at amino acid residues R273 and R248 both result in oncogenic gain-of-function (GOF) phenotypes. Expression of mutant p53 (mtp53) R273H associates with increased cell elasticity, survival under serum deprivation conditions, and increased Poly (ADP ribose) polymerase 1 (PARP1) on the chromatin in the AA-derived TNBC breast cancer cell line MDA-MB-468. We hypothesized that GOF mtp53 R248Q expression could stimulate a similar phenotype in the AA-derived TNBC cell line HCC70. To test this hypothesis we depleted the R248Q protein in the HCC70 cell line using shRNA-mediated knockdown. Using impedance-based real-time analysis we correlated the expression of mtp53 R248Q with increased cell deformability. We also documented that depletion of mtp53 R248Q increased PARP1 in the cytoplasm and decreased PARP1 on the chromatin. We conclude that in the AA-derived TNBC HCC70 cells mtp53 R248Q expression results in a causative tumor associated phenotype. This study supports using the biological markers of high expression of mtp53 R273H or R248Q as additional diagnostics for TNBC resistant subtypes often found in the AA community. Each mtp53 protein must be considered separately and this work adds R248Q to the increasing list of p53 mutations that can be used for diagnostics and drug targeting. Here we report that when R248Q mtp53 proteins are expressed in TNBC, then targeting the gain-of-function pathways may improve treatment efficacy.
Keywords	TNBC ; African American ; p53 ; impedance ; deformability ; Medicine ; R
Subject code	616
Language	English
Publishing date	2015-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Impedimetric detection of mutant p53 biomarker-driven metastatic breast cancers under hyposmotic pressure.

Shi, Menglu / Shtraizent, Nataly / Polotskaia, Alla / Bargonetti, Jill / Matsui, Hiroshi

PloS one

2014 Volume 9, Issue 6, Page(s) e99351

Abstract: In cancer cells, the oncogenic mutant p53 (mtp53) protein is present at high levels and gain-of-function (GOF) activities with more expression of mtp53 proteins contribute to tumor growth and metastasis. Robust analytical approaches that probe the degree ...

Abstract	In cancer cells, the oncogenic mutant p53 (mtp53) protein is present at high levels and gain-of-function (GOF) activities with more expression of mtp53 proteins contribute to tumor growth and metastasis. Robust analytical approaches that probe the degree of metastasis of cancer cells in connection with the mtp53 activity will be extremely useful not only for establishing a better cancer prognosis but also understanding the fundamental mechanism of mtp53 oncogenic action. Here we assessed the influence of mtp53 in breast cancers to the mechanical property of breast cancer cells. Recently, ovarian and kidney cancer cell lines have been shown to have higher cellular elasticity as compared to normal cells assessed by monitoring the degree of deformation under hyposmotic pressure. To make fast detection in large scale, the impedance measurement was applied to monitor the swelling ratio of cells with time. The results showed that knockdown of mtp53 leads to decrease in cell swelling. In addition, by means of two types of impedimetric detection systems we consistently detected enhancement of impedance signal in mtp53-expressing breast cancer cells. Based on this observation we hypothesize that highly expressed mtp53 in metastatic mutant breast cancers can promote tumor progression by making cells more deformable and easier to spread out through extracellular matrix. The identification via the electric measurement can be accomplished within 10 minutes. All results in this report suggest that electric probing for the extent of the mtp53 expression of breast cancer cells may serve as a meaningful fingerprint for the cancer diagnostics, and this outcome will also have an important clinical implication for the development of mtp53-based targeting for tumor detection and treatment.
MeSH term(s)	Biomarkers, Tumor/genetics ; Biomarkers, Tumor/metabolism ; Biomechanical Phenomena ; Breast Neoplasms/pathology ; Cell Line, Tumor ; Cell Membrane/physiology ; Cell Size ; Elasticity ; Female ; Humans ; Neoplasm Metastasis ; Osmotic Pressure ; Tumor Suppressor Protein p53/genetics ; Tumor Suppressor Protein p53/metabolism
Chemical Substances	Biomarkers, Tumor ; TP53 protein, human ; Tumor Suppressor Protein p53
Language	English
Publishing date	2014-06-17
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0099351
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