Group 1: Trauma Operation for Otherwise Healthy Patients Clinical Trial
— RING UB LIGASEOfficial title:
The Role of RING Ubiquitin Ligases in Biologic and Oncologic Processes in Tissues of Mesenchymal Origin
| Verified date | February 2015 |
| Source | Rambam Health Care Campus |
| Contact | n/a |
| Is FDA regulated | No |
| Health authority | Israel: Ethics Commission |
| Study type | Observational |
Stage 1:
Bone marrow will be collected from otherwise healthy patients undergoing orthopedic surgery
- arthroplasty for the treatment of degenerative joint disease or traumatised patients. The
bone marrow will be collected from disposable tissue that is removed during the normal
sequence of the surgery. These cells will be used for:
Establishment of the role of RING ubiquitin ligases in osteogenic progenitors proliferation
and differentiation in culture.
I. To determine the levels of RING ubiquitin ligases mRNA and protein in differentiating
human mesenchymal precursor cells: To test the correlation between levels of RING ubiquitin
ligases and degree of osteogenic differentiation, we will extract mesenchymal precursor
cells from the collected tissue and test the mRNA protein level of RING ubiquitin ligases
upon induction of differentiation. Specifically, we will compare the levels of RING
ubiquitin ligases before and after the initiation of differentiational stimulus in time
dependent manner by western blot and real time PCR analysis.
II. To test the impact of specific RING ubiquitin ligases on differentiation of human
mesenchymal precursor cells: to test the impact of RING ubiquitin ligases on the
differentiation of mesenchymal precursor cell from the collected tissue by testing the
expression of classical differentiational bone markers by flow cytometry (fibronectin,
CD105) and by Alkaline phosphatase (ALP) activity assay. For this aim we developed both a
constitutive and Dox-regulated conditional overexpression and shRNA lenti-viral systems that
enables efficient modulation of these RING ubiquitin ligases level.
III. Determine the role of RING ubiquitin ligases in proliferation and survival of human
mesenchymal precursor cells: Via inhibition or overexpression of ligases in mesenchymal
progenitor cells we will test the role of these ligases in proliferation and survival of
mesenchymal precursors by using MTT assay, Propidion-Iodid (PI) and tunnel assays in flow
cytometry analysis.
Stage 2:
Collection of connective tissue from patients with malignancies of musculoskeletal origin.
The tissue that will be used is part of the resected tumor specimens. The tissue will be
used for:
The establishment of the role/s of RING ligases in musculoskeletal cancers using cell
culture and in vivo activation. Test if the expression of selected positive candidates from
stage 1 correlates with cancer development and progression in human-derived samples
Independently of our mechanistic experiment we aim to determine the relevance of these
ligases to human musculoskeletal cancers. As the first step we will screen primary tumor
biopsies at the protein level correlates with cancer grade and prognosis. Toward this aim we
recently generated a highly specific several anti-monoclonal antibodies in our laboratory as
well use comercial available antibodies .
| Status | Recruiting |
| Enrollment | 90 |
| Est. completion date | July 2019 |
| Est. primary completion date | July 2019 |
| Accepts healthy volunteers | No |
| Gender | Both |
| Age group | 18 Years and older |
| Eligibility |
Group 1: - Inclusion Criteria: - Healthy patients undergoing orthopedic surgery - arthroplasty for the treatment of degenerative joint disease or traumatised patients. - Exclusion Criteria: - Any malignancy in the past or active infection. Group 2: - Inclusion Criteria: - patients with malignancies of musculoskeletal origin. - Exclusion Criteria: - Any malignancy of unknown origin or malignancy of multiple origins or active infection. |
Observational Model: Cohort
| Country | Name | City | State |
|---|---|---|---|
| Israel | Rambam MC | Haifa |
| Lead Sponsor | Collaborator |
|---|---|
| Rambam Health Care Campus | Technion, Israel Institute of Technology |
Israel,
| Type | Measure | Description | Time frame | Safety issue |
|---|---|---|---|---|
| Primary | Protein and mRNA levels of RING ubiquitin ligases during osteogenesis | To determine the levels of RING ubiquitin ligases mRNA and protein in differentiating human mesenchymal precursor cells. We will extract mesenchymal precursor cells from the collected tissue and test the mRNA and protein levels of RING ubiquitin ligases upon induction of differentiation. | 5 years | No |
| Primary | Protein and mRNA levels of RING ubiquitin ligases in musculoskeletal tumors and its correlation to staging and grading | It is not clear if RING ubiquitin ligases protein levels are elevated in soft tissue-derived cancers (primary and metastatic tumors) and whether there is a correlation between RING ubiquitin ligases levels and staging and grading of soft tissue-derived cancers. For this reason, we will check whether RING ubiquitin ligases expression correlates with cancer development and progression in human-derived samples. We will determine the relevance of RING ubiquitin ligases to human musculoskeletal cancers. To do this we will screen primary tumor biopsies for RNA and protein levels of RING ubiquitin ligases . | Time Frame: * 5 years | No |