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Article ; Online: Potential Use of College of American Pathologists Accredited Biorepositories to Bridge Unmet Need for Medical Refrigeration Using Ultralow Temperature Storage for COVID-19 Vaccine or Drug Storage.

Kaushal, Sharmeela / Molinolo, Alfredo

2021 Volume 19, Issue 3, Page(s) 154–155

MeSH term(s)	COVID-19/epidemiology ; COVID-19/prevention & control ; COVID-19 Vaccines ; Drug Storage ; Humans ; Refrigeration ; SARS-CoV-2 ; United States
Chemical Substances	COVID-19 Vaccines
Language	English
Publishing date	2021-04-29
Publishing country	United States
Document type	Letter
ZDB-ID	2593993-2
ISSN	1947-5543 ; 1947-5535
ISSN (online)	1947-5543
ISSN	1947-5535
DOI	10.1089/bio.2020.0163
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Article ; Online: Immunohistochemical Techniques for Phosphoproteins.

Sanz Ressel, Berenice L / Molinolo, Alfredo A

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

2022 Volume 2593, Page(s) 259–264

Abstract: The use of immunohistochemical techniques to study the patterns of protein phosphorylation has revolutionized the study of signaling pathways. This technique allows detecting the phosphorylated state of signaling proteins in formalin-fixed and paraffin- ... ...

Abstract	The use of immunohistochemical techniques to study the patterns of protein phosphorylation has revolutionized the study of signaling pathways. This technique allows detecting the phosphorylated state of signaling proteins in formalin-fixed and paraffin-embedded tissue sections by using phosphospecific antibodies. This chapter describes in detail the immunohistocshemical protocols from which the study of phosphoproteins in tissue sections can be approached.
MeSH term(s)	Paraffin Embedding ; Phosphoproteins ; Immunohistochemistry ; Formaldehyde
Chemical Substances	Phosphoproteins ; Formaldehyde (1HG84L3525)
Language	English
Publishing date	2022-12-13
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2811-9_17
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Article: RNF185 control of COL3A1 expression limits prostate cancer migration and metastatic potential.

Van Espen, Benjamin / Oo, Htoo Zarni / Collins, Colin / Fazli, Ladan / Molinolo, Alfredo / Murad, Rabi / Gleave, Martin / Ronai, Ze'ev A

bioRxiv : the preprint server for biology

2023

Abstract: RNF185 is a RING finger domain-containing ubiquitin ligase implicated in ER-associated degradation. Prostate tumor patient data analysis revealed a negative correlation between RNF185 expression and prostate cancer progression and metastasis. Likewise, ... ...

Abstract	RNF185 is a RING finger domain-containing ubiquitin ligase implicated in ER-associated degradation. Prostate tumor patient data analysis revealed a negative correlation between RNF185 expression and prostate cancer progression and metastasis. Likewise, several prostate cancer cell lines exhibited greater migration and invasion capabilities in culture upon RNF185 depletion. Subcutaneous inoculation of mouse prostate cancer MPC3 cells stably expressing shRNA against RNF185 into mice resulted in larger tumors and more frequent lung metastases. RNA-sequencing and Ingenuity Pathway Analysis identified wound healing and cellular movement among the most significant pathways upregulated in RNF185-depleted, compared to control prostate cancer cells. Gene Set Enrichment Analyses performed in samples from patients harboring low RNF185 expression and in RNF185-depleted lines confirmed the deregulation of genes implicated in EMT. Among those, COL3A1 was identified as the primary mediator of RNF185's ability to impact migration phenotypes. Correspondingly, enhanced migration and metastasis of RNF185 KD prostate cancer cells were attenuated upon co-inhibition of COL3A1. Our results identify RNF185 as a gatekeeper of prostate cancer metastasis, partly
Language	English
Publishing date	2023-07-01
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.06.29.547118
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Article ; Online: Ubiquitin Ligases Siah1a/2 Control Alveolar Macrophage Functions to Limit Carcinogen-Induced Lung Adenocarcinoma.

Scortegagna, Marzia / Du, Yuanning / Bradley, Linda M / Wang, Kun / Molinolo, Alfredo / Ruppin, Eytan / Murad, Rabi / Ronai, Ze'ev A

Cancer research

2023 Volume 83, Issue 12, Page(s) 2016–2033

Abstract: Cellular components of the tumor microenvironment, including myeloid cells, play important roles in the progression of lung adenocarcinoma (LUAD) and its response to therapy. Here, we characterize the function of the ubiquitin ligases Siah1a/2 in ... ...

Abstract	Cellular components of the tumor microenvironment, including myeloid cells, play important roles in the progression of lung adenocarcinoma (LUAD) and its response to therapy. Here, we characterize the function of the ubiquitin ligases Siah1a/2 in regulating the differentiation and activity of alveolar macrophages (AM) and assess the implication of Siah1a/2 control of AMs for carcinogen-induced LUAD. Macrophage-specific genetic ablation of Siah1a/2 promoted accumulation of AMs with an immature phenotype and increased expression of protumorigenic and pro-inflammatory Stat3 and β-catenin gene signatures. Administration of urethane to wild-type mice promoted enrichment of immature-like AMs and lung tumor development, which was enhanced by macrophage-specific Siah1a/2 ablation. The profibrotic gene signature seen in Siah1a/2-ablated immature-like macrophages was associated with increased tumor infiltration of CD14+ myeloid cells and poorer survival of patients with LUAD. Single-cell RNA-seq confirmed the presence of a cluster of immature-like AMs expressing a profibrotic signature in lungs of patients with LUAD, a signature enhanced in smokers. These findings identify Siah1a/2 in AMs as gatekeepers of lung cancer development. Significance: The ubiquitin ligases Siah1a/2 control proinflammatory signaling, differentiation, and profibrotic phenotypes of alveolar macrophages to suppress lung carcinogenesis.
MeSH term(s)	Animals ; Mice ; Macrophages, Alveolar/enzymology ; Macrophages, Alveolar/immunology ; Adenocarcinoma of Lung/chemically induced ; Lung Neoplasms/chemically induced ; Tumor Microenvironment ; Ubiquitin-Protein Ligases/metabolism ; Male ; Mice, Inbred C57BL ; Mice, Knockout
Chemical Substances	Siah1a protein, mouse (EC 2.3.2.27) ; Siah2 protein, mouse (EC 2.3.2.27) ; Ubiquitin-Protein Ligases (EC 2.3.2.27)
Language	English
Publishing date	2023-04-20
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1432-1
ISSN	1538-7445 ; 0008-5472
ISSN (online)	1538-7445
ISSN	0008-5472
DOI	10.1158/0008-5472.CAN-23-0258
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Article ; Online: Correction: a phase 1b study of zilovertamab in combination with paclitaxel for locally advanced/unresectable or metastatic HER2-negative breast cancer.

Shatsky, Rebecca A / Batra-Sharma, Hemali / Helsten, Teresa / Schwab, Richard B / Pittman, Emily I / Pu, Minya / Weihe, Elizabeth / Ghia, Emanuela M / Rassenti, Laura Z / Molinolo, Alfredo / Cabrera, Betty / Breitmeyer, James B / Widhopf Ii, George F / Messer, Karen / Jamieson, Catriona / Kipps, Thomas J / Parker, Barbara A

Breast cancer research : BCR

2024 Volume 26, Issue 1, Page(s) 46

Language	English
Publishing date	2024-03-13
Publishing country	England
Document type	Published Erratum
ZDB-ID	2015059-3
ISSN	1465-542X ; 1465-5411
ISSN (online)	1465-542X
ISSN	1465-5411
DOI	10.1186/s13058-024-01805-w
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Article ; Online: Vulnerabilities of Cancer Patients and Their Effects on Informed Consent for Biobanking.

Kyle, Mason / Cortez, Diana / Carbonell, Blaze / Masmila, Edgar / Molinolo, Alfredo / Kaushal, Sharmeela

Biopreservation and biobanking

2021 Volume 19, Issue 6, Page(s) 543–548

MeSH term(s)	Biological Specimen Banks ; Humans ; Informed Consent ; Neoplasms
Language	English
Publishing date	2021-08-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	2593993-2
ISSN	1947-5543 ; 1947-5535
ISSN (online)	1947-5543
ISSN	1947-5535
DOI	10.1089/bio.2020.0156
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Article ; Online: Real-Time Temperature Mapping in Ultra-Low Freezers as a Standard Quality Assessment.

Powell, Samantha / Molinolo, Alfredo / Masmila, Edgar / Kaushal, Sharmeela

Biopreservation and biobanking

2019 Volume 17, Issue 2, Page(s) 139–142

Abstract: Adequate preservation of biospecimens has been proven to be critical to obtain reliable and reproducible results in genomics, transcriptomics, proteomics, and many other assays. Most biological assays can be performed on specimens preserved in -80°C ... ...

Abstract	Adequate preservation of biospecimens has been proven to be critical to obtain reliable and reproducible results in genomics, transcriptomics, proteomics, and many other assays. Most biological assays can be performed on specimens preserved in -80°C ultra-low freezers, but their quality can be influenced by temperature variability within storage units. Thus, regulatory standards such as those from the College of American Pathologists (CAP), the federal Clinical Laboratory Improvement Amendments, and standards from the Food and Drug Administration require temperature mapping, a standard quality assessment for accreditation when using ultra-low freezers for long-term biospecimen storage. The current mapping methods, providing annual/periodic data, may not be adequate indicators of temperature stability within the different zones of the freezers. In addition, they frequently require manual handling of biospecimens periodically, as they require freezers to be emptied or rearranged temporarily for the installation of temperature probes, risking the integrity of biospecimen quality. In this article, we describe a novel monitoring methodology based on real-time temperature reading of multiple zones by permanently installing thermocouples. An online cloud-based application records temperature variations within 1 minute intervals, and its 24/7 alert system triggers text alarm messages to a predefined set of users when temperature values increase above preset defaults. This provides an opportunity to take remedial action and to obtain a better-quality assessment. Our results indicate that real-time temperature monitoring at multiple zones of a freezer with a 1 minute resolution is a stable and sustainable methodology and, most importantly, lowers the risk of compromising the quality of the biospecimen. The design and use of the real-time monitoring system for ultra-low freezers is one of the acceptable methods by CAP to ensure the stability of biospecimen quality during long-term storage.
MeSH term(s)	Cryopreservation/instrumentation ; Cryopreservation/methods ; Cryopreservation/standards ; Quality Control ; Specimen Handling/instrumentation ; Specimen Handling/methods ; Specimen Handling/standards ; Thermography
Language	English
Publishing date	2019-03-26
Publishing country	United States
Document type	Journal Article
ZDB-ID	2593993-2
ISSN	1947-5543 ; 1947-5535
ISSN (online)	1947-5543
ISSN	1947-5535
DOI	10.1089/bio.2018.0108
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Article ; Online: The molecular underpinning of geminin-overexpressing triple-negative breast cancer cells homing specifically to lungs.

Sami, Eman / Bogan, Danielle / Molinolo, Alfredo / Koziol, Jim / ElShamy, Wael M

Cancer gene therapy

2021 Volume 29, Issue 3-4, Page(s) 304–325

Abstract: Triple-negative breast cancer (TNBCs) display lung metastasis tropism. However, the mechanisms underlying this organ-specific pattern remains to be elucidated. We sought to evaluate the utility of blocking extravasation to prevent lung metastasis. To ... ...

Abstract	Triple-negative breast cancer (TNBCs) display lung metastasis tropism. However, the mechanisms underlying this organ-specific pattern remains to be elucidated. We sought to evaluate the utility of blocking extravasation to prevent lung metastasis. To identify potential geminin overexpression-controlled genetic drivers that promote TNBC tumor homing to lungs, we used the differential/suppression subtractive chain (D/SSC) technique. A geminin overexpression-induced lung metastasis gene signature consists of 24 genes was discovered. We validated overexpression of five of these genes (LGR5, HAS2, CDH11, NCAM2, and DSC2) in worsening lung metastasis-free survival in TNBC patients. Our data demonstrate that LGR5-induced β-catenin signaling and stemness in TNBC cells are geminin-overexpression dependent. They also demonstrate for the first-time expression of RSPO2 in mouse lung tissue only and exacerbation of its secretion in the circulation of mice that develop geminin overexpressing/LGR5
MeSH term(s)	Animals ; Breast/metabolism ; Breast/pathology ; Cell Line, Tumor ; Desmocollins ; Geminin/metabolism ; Humans ; Lung/pathology ; Lung Neoplasms/diagnosis ; Lung Neoplasms/genetics ; Mice ; Neural Cell Adhesion Molecules ; Triple Negative Breast Neoplasms/metabolism
Chemical Substances	Desmocollins ; Dsc2 protein, mouse ; Geminin ; NCAM2 protein, human ; Ncam2 protein, mouse ; Neural Cell Adhesion Molecules
Language	English
Publishing date	2021-03-15
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1212513-1
ISSN	1476-5500 ; 0929-1903
ISSN (online)	1476-5500
ISSN	0929-1903
DOI	10.1038/s41417-021-00311-x
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Zs.A 4125: Show issues

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Article ; Online: A phase 1b study of zilovertamab in combination with paclitaxel for locally advanced/unresectable or metastatic Her2-negative breast cancer.

Shatsky, Rebecca A / Batra-Sharma, Hemali / Helsten, Teresa / Schwab, Richard B / Pittman, Emily I / Pu, Minya / Weihe, Elizabeth / Ghia, Emanuela M / Rassenti, Laura Z / Molinolo, Alfredo / Cabrera, Betty / Breitmeyer, James B / Widhopf, George F / Messer, Karen / Jamieson, Catriona / Kipps, Thomas J / Parker, Barbara A

Breast cancer research : BCR

2024 Volume 26, Issue 1, Page(s) 32

Abstract: Background: Zilovertamab is a humanized monoclonal antibody targeting ROR1, an onco-embryonic antigen expressed by malignant cells of a variety of solid tumors, including breast cancer. A prior phase 1 study showed that zilovertamab was well tolerated ... ...

Abstract	Background: Zilovertamab is a humanized monoclonal antibody targeting ROR1, an onco-embryonic antigen expressed by malignant cells of a variety of solid tumors, including breast cancer. A prior phase 1 study showed that zilovertamab was well tolerated and effective in inhibiting ROR1-signaling, which leads to activation of ERK1/2, NF-κB, and NRF2 target genes. This phase 1b study evaluated the safety and tolerability of zilovertamab with paclitaxel in patients with advanced breast cancer. Patients and methods: Eligible patients had locally advanced, unresectable, or metastatic HER2 Results: Study patients had received a median of 4 prior therapies (endocrine therapy + chemotherapy) for locally advanced, unresectable, or metastatic disease. No patient discontinued therapy due to toxicity ascribed to zilovertamab. Adverse events were consistent with the known safety profile of paclitaxel. Of 16 patients, 6 (38%) had a partial response, and 6/16 (38%) patients had stable disease as best tumor response. Conclusion: The combination of zilovertamab and paclitaxel was safe and well tolerated in heavily pre-treated advanced breast cancer patients. Further evaluation of ROR1 targeting in breast cancer patients with zilovertamab is warranted. Trial registration: NCT02776917. Registered on ClinicalTrials.gov on 05/17/2016.
MeSH term(s)	Humans ; Female ; Breast Neoplasms/drug therapy ; Breast Neoplasms/genetics ; Breast Neoplasms/pathology ; Paclitaxel/therapeutic use ; Receptor, ErbB-2/genetics ; Antibodies, Monoclonal, Humanized ; Antineoplastic Combined Chemotherapy Protocols/adverse effects
Chemical Substances	Paclitaxel (P88XT4IS4D) ; Receptor, ErbB-2 (EC 2.7.10.1) ; Antibodies, Monoclonal, Humanized
Language	English
Publishing date	2024-02-26
Publishing country	England
Document type	Clinical Trial, Phase I ; Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2015059-3
ISSN	1465-542X ; 1465-5411
ISSN (online)	1465-542X
ISSN	1465-5411
DOI	10.1186/s13058-024-01782-0
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Zs.A 5494: Show issues

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Article ; Online: RNF185 Control of COL3A1 Expression Limits Prostate Cancer Migration and Metastatic Potential.

Van Espen, Benjamin / Oo, Htoo Zarni / Collins, Colin / Fazli, Ladan / Molinolo, Alfredo / Yip, Kevin / Murad, Rabi / Gleave, Martin / Ronai, Ze'ev A

Molecular cancer research : MCR

2023 Volume 22, Issue 1, Page(s) 41–54

Abstract	RNF185 is a RING finger domain-containing ubiquitin ligase implicated in ER-associated degradation. Prostate tumor patient data analysis revealed a negative correlation between RNF185 expression and prostate cancer progression and metastasis. Likewise, several prostate cancer cell lines exhibited greater migration and invasion capabilities in culture upon RNF185 depletion. Subcutaneous inoculation of mouse prostate cancer MPC3 cells stably expressing short hairpin RNA against RNF185 into mice resulted in larger tumors and more frequent lung metastases. RNA-sequencing and Ingenuity Pathway Analysis identified wound-healing and cellular movement among the most significant pathways upregulated in RNF185-depleted lines, compared with control prostate cancer cells. Gene Set Enrichment Analyses performed in samples from patients harboring low RNF185 expression and in RNF185-depleted lines confirmed the deregulation of genes implicated in epithelial-to-mesenchymal transition. Among those, COL3A1 was identified as the primary mediator of RNF185's ability to impact migration phenotypes. Correspondingly, enhanced migration and metastasis of RNF185 knockdown (KD) prostate cancer cells were attenuated upon co-inhibition of COL3A1. Our results identify RNF185 as a gatekeeper of prostate cancer metastasis, partly via its control of COL3A1 availability. Implications: RNF185 is identified as an important regulator of prostate cancer migration and metastasis, in part due to its regulation of COL3A1. Both RNF185 and COL3A1 may serve as novel markers for prostate tumors.
MeSH term(s)	Male ; Humans ; Mice ; Animals ; Prostatic Neoplasms/pathology ; Prostate/pathology ; Cell Movement/genetics ; Epithelial-Mesenchymal Transition ; RNA, Small Interfering ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Collagen Type III/genetics ; Collagen Type III/metabolism ; Mitochondrial Proteins/genetics ; Ubiquitin-Protein Ligases/genetics ; Ubiquitin-Protein Ligases/metabolism
Chemical Substances	RNA, Small Interfering ; COL3A1 protein, human ; Collagen Type III ; RNF185 protein, human (EC 2.3.2.27) ; Mitochondrial Proteins ; Ubiquitin-Protein Ligases (EC 2.3.2.27)
Language	English
Publishing date	2023-10-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	2098788-2
ISSN	1557-3125 ; 1541-7786
ISSN (online)	1557-3125
ISSN	1541-7786
DOI	10.1158/1541-7786.MCR-23-0512
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