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Donald Kufe - One of the best experts on this subject based on the ideXlab platform.
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Mucin 1 c terminal subunit oncoprotein is a target for small molecule inhibitors
Molecular Pharmacology, 2011Co-Authors: Yongchun Zhou, Hasan Rajabi, Donald KufeAbstract:Mucin 1 (MUC1) is a heterodimeric protein that is overexpressed in diverse human carcinomas. The oncogenic function of the MUC1 C-terminal subunit (MUC1-C) subunit is dependent on the formation of dimers through its cytoplasmic domain; however, it is not known whether MUC1-C can be targeted with small-molecule inhibitors. In the present work, an assay using the MUC1-C cytoplasmic domain (MUC1-CD) was established to screen small-molecule libraries for compounds that block its dimerization. Using this approach, the flavone apigenin was identified as an inhibitor of MUC1-CD dimerization in vitro and in cells. By contrast, the structurally related flavone baicalein was ineffective in blocking the formation of MUC1-CD dimers. In concert with these results, apigenin, and not baicalein, blocked the localization of MUC1-C to the nucleus. MUC1-C activates MUC1 gene expression in an autoinductive loop, and apigenin, but not baicalein, treatment was associated with down-regulation of MUC1 mRNA levels and MUC1-C protein. The results also demonstrate that apigenin-induced suppression of MUC1-C expression is associated with apoptotic cell death and loss of clonogenic survival. These findings represent the first demonstration that the MUC1-C cytoplasmic domain is a target for the development of small-molecule inhibitors.
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A monoclonal antibody against the oncogenic Mucin 1 cytoplasmic domain.
Hybridoma (2005), 2011Co-Authors: Govind Panchamoorthy, Hala Rehan, Akriti Kharbanda, Rehan Ahmad, Donald KufeAbstract:Mucin 1 (MUC1) is a heterodimeric protein that is aberrantly overexpressed in diverse human carcinomas and certain hematologic malignancies. The transmembrane MUC1-C subunit confers tumorigenicity and is a target for anti-cancer drug development. In this regard, the MUC1-C cytoplasmic domain interacts with multiple effectors that have been linked to transformation. Here we report on the generation of a mouse monoclonal antibody (MAb) against the human MUC1-C cytoplasmic domain (MUC1-CD). This IgG1 MAb, designated anti-MUC1-CD, reacts with the NYGQLDIFP epitope. We show that anti-MUC1-CD is useful in immunoblotting and immunoprecipitation experiments. In addition, anti-MUC1-CD can be used to detect expression of the MUC1-C subunit in formalin-fixed, paraffin-embedded tissues. The MUC1-C inhibitor has entered Phase I evaluation for patients with refractory solid tumors. The present results indicate that the anti-MUC1-CD antibody could be useful as a biomarker to identify patients with tumors that may be responsive to MUC1-C inhibitors.
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direct targeting of the Mucin 1 oncoprotein blocks survival and tumorigenicity of human breast carcinoma cells
Cancer Research, 2009Co-Authors: Deepak Raina, Rehan Ahmad, Li Yin, Surender Kharbanda, Maya Datt Joshi, Takeshi Kawano, Baldev Vasir, David Avigan, Donald KufeAbstract:The Mucin 1 (MUC1) oncoprotein is aberrantly overexpressed by approximately 90% of human breast cancers. However, there are no effective agents that directly inhibit MUC1 and induce death of breast cancer cells. We have synthesized a MUC1 inhibitor (called GO-201) that binds to the MUC1 cytoplasmic domain and blocks the formation of MUC1 oligomers in cells. GO-201, and not an altered version, attenuates targeting of MUC1 to the nucleus of human breast cancer cells, disrupts redox balance, and activates the DNA damage response. GO-201 also arrests growth and induces necrotic death. By contrast, the MUC1 inhibitor has no effect on cells null for MUC1 expression or nonmalignant mammary epithelial cells. Administration of GO-201 to nude mice bearing human breast tumor xenografts was associated with loss of tumorigenicity and extensive necrosis, which results in prolonged regression of tumor growth. These findings show that targeting the MUC1 oncoprotein is effective in inducing death of human breast cancer cells in vitro and in tumor models.
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Human Mucin 1 oncoprotein represses transcription of the p53 tumor suppressor gene.
Cancer research, 2007Co-Authors: Xiaolong Wei, Donald KufeAbstract:The Mucin 1 (MUC1) heterodimeric protein is aberrantly overexpressed in human breast cancers and induces transformation. The MUC1 COOH-terminal subunit (MUC1-C) is targeted to the nucleus of transformed cells, where it interacts with p53 and regulates p53-mediated transcription. The present studies show that MUC1 represses activation of the p53 gene and that MUC1-C occupies the PE21 element in the p53 proximal promoter. Previous work has shown that the Kruppel-like factor 4 (KLF4) transcription factor represses p53 transcription by binding to the PE21 element. Our results show that MUC1-C binds constitutively to KLF4, occupies PE21 with KLF4, and enhances the KLF4 occupancy of PE21. The results also show that MUC1-C increases the recruitment of histone deacetylases 1/3, deacetylation of core histones, and repression of p53 transcription. These findings indicate that overexpression of MUC1, as found in human breast cancer cells, is of functional importance to repression of the p53 gene.
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Mucin 1 Oncoprotein Blocks Hypoxia-inducible Factor 1α Activation in a Survival Response to Hypoxia
The Journal of biological chemistry, 2006Co-Authors: Li Yin, Surender Kharbanda, Donald KufeAbstract:Resistance of carcinoma cells to hypoxic stress is of importance to the growth of solid tumors. The Mucin 1 (MUC1) oncoprotein is aberrantly overexpressed by most human carcinomas; however, there is no known relationship between MUC1 and the hypoxic stress response. The present work has demonstrated that MUC1 attenuates activation of hypoxia-inducible factor-1α (HIF-1α), a regulator of gene transcription in the response of cells to hypoxic stress. In cells with stable gain and loss of MUC1 function, we have shown that MUC1 up-regulates prolyl hydroxylase 3 (PHD3) expression and promotes HIF-1α degradation. PHD activity is attenuated by increases in reactive oxygen species (ROS) generated in the hypoxic stress response. Our results further demonstrate that MUC1 blocks hypoxia-induced increases in ROS and thereby potentiates PHD-mediated HIF-1α suppression. Importantly, MUC1 also blocks hypoxia-induced apoptosis and necrosis by suppressing accumulation of ROS. These findings indicate that MUC1 attenuates HIF-1α activation in a survival response to hypoxic stress.
Mats Ohlin - One of the best experts on this subject based on the ideXlab platform.
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Antigens for the selection of pan-variable number of tandem repeats motif-specific human antibodies against Mucin-1.
Journal of immunological methods, 2006Co-Authors: Jonas Persson, Mats OhlinAbstract:Epitopes found on Mucin-1 are differentially expressed on tumour versus normal tissue. Such epitopes have also been shown to have a potential in immunotherapy and tumour detection. The major epitope explored in this context is located within the variable number of tandem repeats. It has however recently been demonstrated that this epitope exists in several sequence variants. The standard sequence is highly antigenic while the most common sequence variant is much less so. We have now explored routes employing defined synthetic antigens to ensure the development of human recombinant antibody specificities targeting both sequence variants of this epitope. These antibodies may serve as a platform for the development of human antibodies for efficient targeting of Mucin-1 in human disease.
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Research paper Antigens for the selection of pan-variable number of tandem repeats motif-specific human antibodies against Mucin-1
2006Co-Authors: Jonas Persson, Mats OhlinAbstract:Epitopes found on Mucin-1 are differentially expressed on tumour versus normal tissue. Such epitopes have also been shown to have a potential in immunotherapy and tumour detection. The major epitope explored in this context is located within the variable number of tandem repeats. It has however recently been demonstrated that this epitope exists in several sequence variants. The standard sequence is highly antigenic while the most common sequence variant is much less so. We have now explored routes employing defined synthetic antigens to ensure the development of human recombinant antibody specificities targeting both sequence variants of this epitope. These antibodies may serve as a platform for the development of human antibodies for efficient targeting of Mucin-1 in human disease. © 2006 Elsevier B.V. All rights reserved.
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Antibody evolution from the centre to the periphery: applied to a human antibody fragment recognising the tumour-associated antigen Mucin-1.
Journal of molecular biology, 2002Co-Authors: Pernilla Jirholt, Carl A.k. Borrebaeck, Mats OhlinAbstract:Mucin-1 has proven to be a suitable target for antibody-based diagnosis and therapy of certain tumours, but no appropriate human antibody or antibody fragment displaying slow dissociation rate kinetics against this target is available. Since a rapid dissociation character prevents an antibody fragment from remaining at the site of the antigen, this fact may prevent the successful application of a human Mucin-1 specific antibody in diagnosis and therapy. We have now used iterative antibody libraries to evolve a human antibody fragment originally obtained from a naive antibody library. A strategy was devised whereby molecular variants displaying slow dissociation kinetics against the repetitive Mucin-1 tumour-associated antigen can be selected in vitro. The evolved clones, that allowed for a reduced dissociation from the tumour antigen, carried substitutions in the outer parts of the binding site. This demonstrated the ability of this in vitro evolution technique to mimic the process whereby antibodies evolve in vivo. We have thus devised a strategy through which molecular variants displaying slow dissociation from a repetitive target like the Mucin-1 tumour-associated antigen can be obtained in vitro. These or related molecules obtained by this approach will serve as a starting point for the development of fully human antibodies for use in Mucin-1 specific tumour therapy of diagnosis.
Gerard M. J. Bos - One of the best experts on this subject based on the ideXlab platform.
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Expression of aberrantly glycosylated Mucin-1 in ovarian cancer.
Histopathology, 2010Co-Authors: Catharina H. M. J. Van Elssen, Peter W. H. Frings, F.j. Bot, Koen Van De Vijver, Mariska B. Huls, Bob Meek, Pierre Hupperets, Wilfred T. V. Germeraad, Gerard M. J. BosAbstract:Van Elssen C H M J, Frings P W H, Bot F J, Van de Vijver K K, Huls M B, Meek B, Hupperets P, Germeraad W T V & Bos G M J (2010) Histopathology57, 597–606 Expression of aberrantly glycosylated Mucin-1 in ovarian cancer Aims: Mucin 1 (MUC1) is an important tumour-associated antigen (TAA), both overexpressed and aberrantly glycosylated in adenocarcinomas. The aim of this study was to examine the MUC1-glycosylation status of primary ovarian adenocarcinomas and metastatic lesions. Methods and results: Paraffin-embedded tissue sections of 37 primary ovarian adenocarcinomas representing all histotypes (22 serous, five Mucinous, two clear-cell, eight endometrioid), four serous borderline tumours with intraepithelial carcinoma, seven sections of ovarian endometriosis and 13 metastatic lesions were analysed by immunohistochemistry. Non-neoplastic ovarian surface epithelium and serous cystadenomas were used as controls. All epithelia expressed MUC1 protein. Of primary tumours, 76% expressed the differentiation-dependent glycoform and 84% the cancer-associated glycoform (Tn/Sialyl-Tn-epitopes). In metastatic lesions this was 77% and 85%, respectively. Notably, in 57% of ovarian endometriosis and 75% of intraepithelial lesions, the cancer-associated MUC1 epitopes were expressed, whereas normal ovarian surface epithelium and serous cystadenomas did not express these epitopes. Conclusions: The underglycosylated MUC1 epitopes are expressed by all histotypes of primary ovarian adenocarcinomas, by the vast majority of metastatic lesions and by possible ovarian cancer precursor lesions, but not by normal ovarian tissue. These results indicate that MUC1-associated Tn/STn-epitopes are important targets for immunotherapy and diagnostic imaging in ovarian cancer patients.
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Expression of aberrantly glycosylated Mucin-1 in ovarian cancer
Histopathology, 2010Co-Authors: Catharina H. M. J. Van Elssen, Peter W. H. Frings, F.j. Bot, Mariska B. Huls, Bob Meek, Pierre Hupperets, Wilfred T. V. Germeraad, Koen K Van De Vijver, Gerard M. J. BosAbstract:Aims: Mucin 1 (MUC1) is an important tumour-associated antigen (TAA), both over-expressed and aberrantly glycosylated in adenocarcinomas. The objective of this study was to examine the MUC1-glycosylation status in primary ovarian adenocarcinomas and metastatic lesions. Methods and results: Paraffin-embedded tissue sections of 37 primary ovarian adenocarcinomas representing all histotypes (22 serous, 5 Mucinous, 2 clear cell, 8 endometrioid), 4 serous borderline tumours with intraepithelial carcinoma, 7 sections of ovarian endometriosis and 13 metastatic lesions were analysed by immuno-histochemistry. Non-neoplastic ovarian surface epithelium and serous cystadenomas were used as controls. All epithelia expressed MUC1 protein. Of primary tumours, 76% expressed the differentiation-dependent glycoform and 84% the cancer-associated glycoform (Tn/Sialyl-Tn-epitopes). In metastatic lesions this was 77% and 85%, respectively. Notably, in 57% of ovarian endometriosis and 75% of intraepithelial lesions, the cancer-associated MUC1 epitopes were expressed, whereas normal ovarian surface epithelium and serous cystadenomas did not express these epitopes. Conclusions: The underglycosylated MUC1 epitopes are expressed by all histotypes of primary ovarian adenocarcinomas, by the vast majority of metastatic lesions and by ovarian cancer precursor lesions, whereas not by normal ovarian tissue. These results indicate that MUC1-associated Tn/STn-epitopes are important targets for immunotherapy and diagnostic imaging in ovarian cancer patients.
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Surface Mucin-1 does not play a role in dendritic cell migration.
Molecular immunology, 2008Co-Authors: Silvie Cloosen, Mariska B. Huls, Wilfred T. V. Germeraad, Birgit L. M. G. Senden-gijsbers, Jean-hubert Caberg, Joris Vanderlocht, Patrick Roncarati, Pascale Hubert, Philippe Delvenne, Gerard M. J. BosAbstract:Abstract Mucin-1 (MUC1) is a transmembrane glycoprotein that is upregulated upon maturation of dendritic cells (DC) in vitro or in vivo. One of the proposed functions of surface expressed MUC1 is its involvement in migration of cells. We hypothesized that MUC1 is involved in DC migration since mature DC (mDC) are highly migratory cells and MUC1 is upregulated on the surface of DC upon maturation. In this study we cultured DC using two maturation cocktails, one cocktail containing IL-4, GM-CSF, TNFα, PGE2, IL-1β and IL-6 (TP1,6-DC) and the other IL-13, GM-CSF, Ribomunyl and IFN-γ (RI-DC). Both maturation cocktails render DC with a similar surface phenotype including CCR7 expression, but only the former induces a migratory capacity of DC to a CCL19 gradient. To analyze the role of surface-expression of MUC1 on TP1,6-DC, that are capable of migration, expression of MUC1 was prevented by adding an anti-MUC1 antibody (Ab) during the maturation process. Compared with matured DC in the absence of the Ab, no difference was observed in chemokine-induced migratory behaviour between the MUC1+ and MUC1− DC populations in a standard Transwell chemotaxis assay, nor in organotypic cultures. Our data clearly demonstrate that surface MUC1 on DC does not influence intrinsic cell-motility, nor is it involved in cell–cell and cell–matrix dependent migration.
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Mucin-1 is expressed on dendritic cells, both in vitro and in vivo
International immunology, 2004Co-Authors: Silvie Cloosen, Wilfred T. V. Germeraad, Ellen B. M. Van Leeuwen, Birgit L. M. G. Senden-gijsbers, Ellis B. H. Oving, Marco Thio, Ariane Vanclée, Gerard M. J. BosAbstract:Dendritic cells (DCs) are the best professional antigen-presenting cells to stimulate cytotoxic as well as T helper cells and are therefore appropriate candidates for establishing immunotherapy. The concept of our vaccination program is to introduce the tumor-associated antigen Mucin-1 (MUC1) into DCs. Analysis of immature and mature DCs—before transducing the antigen MUC1—already demonstrated expression of MUC1 on in vitro monocyte-derived DCs upon maturation. Different culture methods as well as maturation cocktails showed similar results concerning the upregulation of MUC1 expression. Furthermore, we studied the expression of MUC1 on DCs in vivo. No MUC1 expression was found on blood DCs, or on thymic or tonsil DCs. On the other hand, synovial fluid from patients with arthritis contained DCs that were found to express MUC1. This study shows for the first time that the tumor-associated antigen MUC1 is expressed on in vivo DCs. We further show that MUC1 is also expressed on in vitro cultured bone marrow-derived DCs of human MUC1 transgenic mice, supporting the relevance of this mouse model to the human situation. The observation that MUC1 is present on in vivo DCs suggests a functional role, but this physiological function remains to be elucidated.
Heidi L Rehm - One of the best experts on this subject based on the ideXlab platform.
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development and validation of a mass spectrometry based assay for the molecular diagnosis of Mucin 1 kidney disease
The Journal of Molecular Diagnostics, 2016Co-Authors: Brendan Blumenstiel, Matthew Defelice, Ozge Birsoy, Anthony J Bleyer, Stanislav Kmoch, Todd A Carter, Andreas Gnirke, Kendrah Kidd, Heidi L RehmAbstract:Mucin-1 kidney disease, previously described as medullary cystic kidney disease type 1 (MCKD1, OMIM 174000), is an autosomal dominant tubulointerstitial kidney disease recently shown to be caused by a single-base insertion within the variable number tandem repeat region of the MUC1 gene. Because of variable age of disease onset and often subtle signs and symptoms, clinical diagnosis of Mucin-1 kidney disease and differentiation from other forms of hereditary kidney disease have been difficult. The causal insertion resides in a variable number tandem repeat region with high GC content, which has made detection by standard next-generation sequencing impossible to date. The inherently difficult nature of this mutation required an alternative method for routine detection and clinical diagnosis of the disease. We therefore developed and validated a mass spectrometry–based probe extension assay with a series of internal controls to detect the insertion event using 24 previously characterized positive samples from patients with Mucin-1 kidney disease and 24 control samples known to be wild type for the variant. Validation results indicate an accurate and reliable test for clinically establishing the molecular diagnosis of Mucin-1 kidney disease with 100% sensitivity and specificity across 275 tests called.
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development and validation of a mass spectrometry based assay for the molecular diagnosis of Mucin 1 kidney disease
Elsevier, 2016Co-Authors: Brendan Blumenstiel, Matthew Defelice, Ozge Birsoy, Anthony J Bleyer, Stanislav Kmoch, Todd A Carter, Andreas Gnirke, Kendrah Kidd, Heidi L RehmAbstract:opyright a 2016 American Society for Inve his is an open access article under the CC B ttp://dx.doi.org/10.1016/j.jmoldx.2016.03.003 Mucin-1 kidney disease, previously described as medullary cystic kidney disease type 1 (MCKD1, OMIM 174000), is an autosomal dominant tubulointerstitial kidney disease recently shown to be caused by a single-base insertion within the variable number tandem repeat region of the MUC1 gene. Because of variable age of disease onset and often subtle signs and symptoms, clinical diagnosis of Mucin-1 kidney disease and differentiation from other forms of hereditary kidney disease have been difficult. The causal insertion resides in a variable number tandem repeat region with high GC content, which has made detection by standard next-generation sequencing impossible to date. The inherently difficult nature of this mutation required an alternative method for routine detection and clinical diagnosis of the disease. We therefore developed and validated a mass spectrometryebased probe extension assay with a series of internal controls to detect the insertion event using 24 previously characterized positive samples from patients with Mucin-1 kidney disease and 24 control samples known to be wild type for the variant. Validation results indicate an accurate and reliable test for clinically establishing the molecular diagnosis of Mucin-1 kidney disease with 100% sensitivity and specificity across 275 tests called. (J Mol Diagn 2016, 18: 566e571; http://dx.doi.org/10.1016/j.jmoldx.2016.03.003)
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Development and Validation of a Mass Spectrometry–Based Assay for the Molecular Diagnosis of Mucin-1 Kidney Disease
2016Co-Authors: Brendan Blumenstiel, Matthew Defelice, Ozge Birsoy, Anthony J Bleyer, Stanislav Kmoch, Todd A Carter, Andreas Gnirke, Kendrah Kidd, Heidi L Rehm, Lucienne RoncoAbstract:opyright a 2016 American Society for Inve his is an open access article under the CC B ttp://dx.doi.org/10.1016/j.jmoldx.2016.03.003 Mucin-1 kidney disease, previously described as medullary cystic kidney disease type 1 (MCKD1, OMIM 174000), is an autosomal dominant tubulointerstitial kidney disease recently shown to be caused by a single-base insertion within the variable number tandem repeat region of the MUC1 gene. Because of variable age of disease onset and often subtle signs and symptoms, clinical diagnosis of Mucin-1 kidney disease and differentiation from other forms of hereditary kidney disease have been difficult. The causal insertion resides in a variable number tandem repeat region with high GC content, which has made detection by standard next-generation sequencing impossible to date. The inherently difficult nature of this mutation required an alternative method for routine detection and clinical diagnosis of the disease. We therefore developed and validated a mass spectrometryebased probe extension assay with a series of internal controls to detect the insertion event using 24 previously characterized positive samples from patients with Mucin-1 kidney disease and 24 control samples known to be wild type for the variant. Validation results indicate an accurate and reliable test for clinically establishing the molecular diagnosis of Mucin-1 kidney disease with 100% sensitivity and specificity across 275 tests called. (J Mol Diagn 2016, 18: 566e571; http://dx.doi.org/10.1016/j.jmoldx.2016.03.003)
Ozge Birsoy - One of the best experts on this subject based on the ideXlab platform.
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development and validation of a mass spectrometry based assay for the molecular diagnosis of Mucin 1 kidney disease
The Journal of Molecular Diagnostics, 2016Co-Authors: Brendan Blumenstiel, Matthew Defelice, Ozge Birsoy, Anthony J Bleyer, Stanislav Kmoch, Todd A Carter, Andreas Gnirke, Kendrah Kidd, Heidi L RehmAbstract:Mucin-1 kidney disease, previously described as medullary cystic kidney disease type 1 (MCKD1, OMIM 174000), is an autosomal dominant tubulointerstitial kidney disease recently shown to be caused by a single-base insertion within the variable number tandem repeat region of the MUC1 gene. Because of variable age of disease onset and often subtle signs and symptoms, clinical diagnosis of Mucin-1 kidney disease and differentiation from other forms of hereditary kidney disease have been difficult. The causal insertion resides in a variable number tandem repeat region with high GC content, which has made detection by standard next-generation sequencing impossible to date. The inherently difficult nature of this mutation required an alternative method for routine detection and clinical diagnosis of the disease. We therefore developed and validated a mass spectrometry–based probe extension assay with a series of internal controls to detect the insertion event using 24 previously characterized positive samples from patients with Mucin-1 kidney disease and 24 control samples known to be wild type for the variant. Validation results indicate an accurate and reliable test for clinically establishing the molecular diagnosis of Mucin-1 kidney disease with 100% sensitivity and specificity across 275 tests called.
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development and validation of a mass spectrometry based assay for the molecular diagnosis of Mucin 1 kidney disease
Elsevier, 2016Co-Authors: Brendan Blumenstiel, Matthew Defelice, Ozge Birsoy, Anthony J Bleyer, Stanislav Kmoch, Todd A Carter, Andreas Gnirke, Kendrah Kidd, Heidi L RehmAbstract:opyright a 2016 American Society for Inve his is an open access article under the CC B ttp://dx.doi.org/10.1016/j.jmoldx.2016.03.003 Mucin-1 kidney disease, previously described as medullary cystic kidney disease type 1 (MCKD1, OMIM 174000), is an autosomal dominant tubulointerstitial kidney disease recently shown to be caused by a single-base insertion within the variable number tandem repeat region of the MUC1 gene. Because of variable age of disease onset and often subtle signs and symptoms, clinical diagnosis of Mucin-1 kidney disease and differentiation from other forms of hereditary kidney disease have been difficult. The causal insertion resides in a variable number tandem repeat region with high GC content, which has made detection by standard next-generation sequencing impossible to date. The inherently difficult nature of this mutation required an alternative method for routine detection and clinical diagnosis of the disease. We therefore developed and validated a mass spectrometryebased probe extension assay with a series of internal controls to detect the insertion event using 24 previously characterized positive samples from patients with Mucin-1 kidney disease and 24 control samples known to be wild type for the variant. Validation results indicate an accurate and reliable test for clinically establishing the molecular diagnosis of Mucin-1 kidney disease with 100% sensitivity and specificity across 275 tests called. (J Mol Diagn 2016, 18: 566e571; http://dx.doi.org/10.1016/j.jmoldx.2016.03.003)
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Development and Validation of a Mass Spectrometry–Based Assay for the Molecular Diagnosis of Mucin-1 Kidney Disease
2016Co-Authors: Brendan Blumenstiel, Matthew Defelice, Ozge Birsoy, Anthony J Bleyer, Stanislav Kmoch, Todd A Carter, Andreas Gnirke, Kendrah Kidd, Heidi L Rehm, Lucienne RoncoAbstract:opyright a 2016 American Society for Inve his is an open access article under the CC B ttp://dx.doi.org/10.1016/j.jmoldx.2016.03.003 Mucin-1 kidney disease, previously described as medullary cystic kidney disease type 1 (MCKD1, OMIM 174000), is an autosomal dominant tubulointerstitial kidney disease recently shown to be caused by a single-base insertion within the variable number tandem repeat region of the MUC1 gene. Because of variable age of disease onset and often subtle signs and symptoms, clinical diagnosis of Mucin-1 kidney disease and differentiation from other forms of hereditary kidney disease have been difficult. The causal insertion resides in a variable number tandem repeat region with high GC content, which has made detection by standard next-generation sequencing impossible to date. The inherently difficult nature of this mutation required an alternative method for routine detection and clinical diagnosis of the disease. We therefore developed and validated a mass spectrometryebased probe extension assay with a series of internal controls to detect the insertion event using 24 previously characterized positive samples from patients with Mucin-1 kidney disease and 24 control samples known to be wild type for the variant. Validation results indicate an accurate and reliable test for clinically establishing the molecular diagnosis of Mucin-1 kidney disease with 100% sensitivity and specificity across 275 tests called. (J Mol Diagn 2016, 18: 566e571; http://dx.doi.org/10.1016/j.jmoldx.2016.03.003)