Nuclear Myosin VI - a Therapeutic Target in Breast Cancer

NCT03430024 | Unknown

• 1 other identifier: 17/09/621_Myosin VI
• Study Type: observational
• Target: 120
• Locations: 1 country
• Sites: 1

Brief Summary

Gene expression, the transfer of the genetic code into cellular proteins is one of the most fundamental processes in living cells. This process is orchestrated by protein-based molecular machines, called RNA polymerases that read the DNA sequence to generate messenger RNA (mRNA), which is translated by the cellular machinery to make proteins. Our cells have evolved elaborate regulation mechanisms to control these molecular machines and a breakdown in this regulation leads to diseases such as cancer. Recently, molecules called myosins have been discovered in the genetic storage compartment of the cell (the nucleus) where they interact with RNA polymerases to regulate protein production. This is interesting because myosins are usually found outside the nucleus transporting cellular cargo or generating muscle contraction. In breast cancer cells, myosin is abundant and interacts with the oestrogen receptor. The majority of breast cancer in the UK is oestrogen receptor positive and activation of this receptor is an important factor controlling the growth of cancer cells. Oestrogen receptor activation appears to be dependent upon myosin and this research project will investigate how myosins are targeted to specific genes and how they are themselves regulated. This will greatly enhance our understanding of the role of nuclear myosins in oestrogen receptor positive breast cancer and may identify a novel therapeutic target for future drug development.

Conditions

Breast Cancer

Keywords

Myosin VI; Estrogen receptor

Outcome Measures

Primary Outcomes (3)

• Gene expression of Myosin VI and oestrogen receptor targets in tumour tissue.

  RT-qPCR experiments to determine relative expression of Myosin VI and oestrogen receptor targets in tumour derived RNA and compare with control breast tissue.

  24 hours

• Quantification of Myosin VI protein in tumour tissue.

  Western-blot analysis and cell fractionation to determine relative amounts of Myosin VI compared to total protein extracted from 100mg of tumour tissue in nuclear and cytoplasmic compartments and compare with control tissue.

  24 hours.

• Localisation of Myosin VI on the tumour genome

  Chromatin-immunoprecipitation experiments to determine the position of Myosin VI in the tumour genome and compare with control tissue.

  24 hours.

Secondary Outcomes (1)

• Comparison of nuclear Myosin VI and oestrogen receptor localisation between different breast cancer prognostic groups.

  3 months

Study Arms (2)

Cases

We will enrol 100 women who have been newly diagnosed with T2 (\>2cm) palpable invasive breast cancer having primary surgical treatment at Maidstone Hospital. We will exclude all patients with a metabolic disorder, significant co-morbidities and locally advanced or metastatic disease as well as those with a previous history of cancer treatment. We will collect data on tumour size, grade and phenotype as well as ER, progesterone receptor (PR) and Her-2 expression status and patient demographic information. We will investigate the Association of Myosin VI with oestrogen receptor.

Other: Association of Myosin VI with oestrogen receptor

Controls

A cohort of control breast tissue will be obtained from 20 patients undergoing benign surgical breast procedures. For those control patients having reduction mammoplasties the excised tissue will be core biopsied but patients having other types of benign surgery will have an extra core biopsy taken from breast tissue surrounding the lesion being excised.

Other: Association of Myosin VI with oestrogen receptor

Interventions

Association of Myosin VI with oestrogen receptor OTHER

1. Gene expression analysis - RNA will be extracted from 100mg of tissue and then subjected to RT-qPCR to determine the relative expression levels of several genes, including MVI and ER Targets. We will use the same approach to determine which isoforms of MVI are expressed in the tissue. 2. Protein content analysis - Total protein will be extracted from 100 mg of tissue and western-blot analysis will be used to determine the relative amounts of MVI. Moreover, we will use cell fractionation to determine relative amounts of nuclear versus cytoplasmic protein. 3. Genomic analysis - Chromatin-immunoprecipitation (ChIP) experiments we be performed upon 100 mg of tissue sample. Here we will determine the localisation of MVI on the genome within the different tissues.

Cases; Controls

Eligibility Criteria

• Age: 18 Years - 85 Years
• Sex: female
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

Tissue will be obtained from patients diagnosed with early invasive breast cancer undergoing primary surgery. We will enrol 100 women who have been newly diagnosed with T2 (\>2cm) palpable invasive breast cancer having primary surgical treatment at Maidstone Hospital.

You may qualify if:

• Clinical diagnosis of early breast cancer
• Palpable tumour greater than 2cm
• Scheduled for primary surgical treatment

You may not qualify if:

• Locally advanced breast cancer
• Metastatic breast cancer
• Significant co-morbidities (ASA 4 or above)
• Past history of breast cancer

Sponsors & Collaborators

• Maidstone & Tunbridge Wells NHS Trust: lead
• University of Kent: collaborator

Study Sites (1)

Maidstone and Tunbridge Wells NHS Trust

Maidstone, Kent, ME16 9QQ, United Kingdom

RECRUITING

Related Publications (8)

• Vreugde S, Ferrai C, Miluzio A, Hauben E, Marchisio PC, Crippa MP, Bussi M, Biffo S. Nuclear myosin VI enhances RNA polymerase II-dependent transcription. Mol Cell. 2006 Sep 1;23(5):749-55. doi: 10.1016/j.molcel.2006.07.005.

  PMID: 16949370 BACKGROUND

• Su AI, Welsh JB, Sapinoso LM, Kern SG, Dimitrov P, Lapp H, Schultz PG, Powell SM, Moskaluk CA, Frierson HF Jr, Hampton GM. Molecular classification of human carcinomas by use of gene expression signatures. Cancer Res. 2001 Oct 15;61(20):7388-93.

  PMID: 11606367 BACKGROUND

• Dunn TA, Chen S, Faith DA, Hicks JL, Platz EA, Chen Y, Ewing CM, Sauvageot J, Isaacs WB, De Marzo AM, Luo J. A novel role of myosin VI in human prostate cancer. Am J Pathol. 2006 Nov;169(5):1843-54. doi: 10.2353/ajpath.2006.060316.

  PMID: 17071605 BACKGROUND

• Wang H, Wang B, Zhu W, Yang Z. Lentivirus-Mediated Knockdown of Myosin VI Inhibits Cell Proliferation of Breast Cancer Cell. Cancer Biother Radiopharm. 2015 Oct;30(8):330-5. doi: 10.1089/cbr.2014.1759. Epub 2015 Sep 25.

  PMID: 26407123 BACKGROUND

• Yoshida H, Cheng W, Hung J, Montell D, Geisbrecht E, Rosen D, Liu J, Naora H. Lessons from border cell migration in the Drosophila ovary: A role for myosin VI in dissemination of human ovarian cancer. Proc Natl Acad Sci U S A. 2004 May 25;101(21):8144-9. doi: 10.1073/pnas.0400400101. Epub 2004 May 14.

  PMID: 15146066 BACKGROUND

• Spudich G, Chibalina MV, Au JS, Arden SD, Buss F, Kendrick-Jones J. Myosin VI targeting to clathrin-coated structures and dimerization is mediated by binding to Disabled-2 and PtdIns(4,5)P2. Nat Cell Biol. 2007 Feb;9(2):176-83. doi: 10.1038/ncb1531. Epub 2006 Dec 24.

  PMID: 17187061 BACKGROUND

• Morriswood B, Ryzhakov G, Puri C, Arden SD, Roberts R, Dendrou C, Kendrick-Jones J, Buss F. T6BP and NDP52 are myosin VI binding partners with potential roles in cytokine signalling and cell adhesion. J Cell Sci. 2007 Aug 1;120(Pt 15):2574-85. doi: 10.1242/jcs.007005. Epub 2007 Jul 17.

  PMID: 17635994 BACKGROUND

• Naccache SN, Hasson T, Horowitz A. Binding of internalized receptors to the PDZ domain of GIPC/synectin recruits myosin VI to endocytic vesicles. Proc Natl Acad Sci U S A. 2006 Aug 22;103(34):12735-40. doi: 10.1073/pnas.0605317103. Epub 2006 Aug 14.

  PMID: 16908842 BACKGROUND

MeSH Terms

Conditions

Breast Neoplasms; Deafness, Autosomal Recessive 37

Interventions

Estrogen Receptor alpha

Condition Hierarchy (Ancestors)

Neoplasms by Site; Neoplasms; Breast Diseases; Skin Diseases; Skin and Connective Tissue Diseases

Intervention Hierarchy (Ancestors)

Receptors, Estrogen; Receptors, Steroid; Receptors, Cytoplasmic and Nuclear; Proteins; Amino Acids, Peptides, and Proteins

Study Officials

• Hazel Everest

  Maidstone and Tunbridge Wells NHS Trust

  STUDY DIRECTOR

Central Study Contacts

Karina Cox, MD

CONTACT

00 45 1622 22 4111; karina.cox@nhs.net

Chris Toseland, PhD

CONTACT

01227816515; c.toseland@kent.ac.uk

Study Design

• Study Type: observational
• Observational Model: OTHER
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Consultant Breast and Oncoplastic Surgeon

Study Record Dates

• First Submitted: January 11, 2018
• First Posted: February 12, 2018
• Study Start: May 5, 2019
• Primary Completion: December 1, 2020
• Study Completion: December 1, 2020
• Last Updated: November 13, 2019

Record last verified: 2019-11

Data Sharing

• IPD Sharing: Will not share

Locations

GB (1)