TGF-βs in the eye of patients with neovascular age-related macular degeneration
| ISRCTN | ISRCTN60434145 |
|---|---|
| DOI | https://doi.org/10.1186/ISRCTN60434145 |
| Secondary identifying numbers | AMD001.30 |
- Submission date
- 08/01/2018
- Registration date
- 12/01/2018
- Last edited
- 10/12/2018
- Recruitment status
- No longer recruiting
- Overall study status
- Completed
- Condition category
- Eye Diseases
Plain English summary of protocol
Background and study aim
Wet age-related macular degeneration (AMD) is the commonest cause of blindness in developed countries. Many proteins (called cytokines) are thought to be responsible for this disease. One of these (VEGF-A) is the target of the current therapy. With this study, we want to clarify the role of the cytokines called TGF-beta in the disease to better understand the causes and to develop a more effective therapy.
Who can participate?
Patients aged 50 and over with AMD and cataract patients as controls.
What does the study involve?
Participants affected by AMD receive, as standard treatment, a loading dose of three injections over a three month period. Before each treatment a small sample of aqueous humor (the eyeball is filled with this liquid) is collected by a syringe from the anterior chamber of the eye. Cataract controls are subjected to sample collection just before the planned cataract surgery.
What are the possible benefits and risks of participating?
Study participation may not directly help patients but the information obtained from the study may help to improve the knowledge of the disease and consequently the treatment of people with wet AMD in the future. This is a study embedded in the routine clinical procedures without any additional treatment (except the sample collection), therefore there should be no risks or disadvantages to patients taking part. All participants receive the same treatment as prescribed by their physicians.
Where is the study run from?
Ophthalmology Unit of the Department of Medicine, Surgery and Neuroscience, Siena University Hospital (Italy)
When is the study starting and how long is it expected to run for?
June 2016 to December 2016
Who is funding the study?
University of Siena
Who is the main contact?
Prof. Gian Marco Tosi
gmtosi18@gmail.com
Contact information
Scientific
Ophthalmology Unit
Department of Medicine, Surgery and Neuroscience
University of Siena
Viale Bracci, 1
Siena
53100
Italy
| Phone | +39 0577 586162 |
|---|---|
| gmtosi18@gmail.com |
Study information
| Study design | Observational single-centre case-control study |
|---|---|
| Primary study design | Observational |
| Secondary study design | Case-control study |
| Study setting(s) | Hospital |
| Study type | Other |
| Participant information sheet | Not available in web format, please use the contact details below to request a patient information sheet |
| Scientific title | TGF-β concentrations and activity in the aqueous humor of patients with neovascular age-related macular degeneration before and after ranibizumab treatment |
| Study objectives | The aim of this study is to evaluate the protein concentration of active TGF-β1, TGF-β2, and TGF-β3 in the aqueous humor of patients affected by naïve nAMD, at baseline and after intravitreal anti-VEGF-A injection by performing a luciferase-based reporter assay to test the TGF-β pathway activation by aqueous humor samples of the same patients, with the aim to clarify the role of the TGF-β family members in CNV. |
| Ethics approval(s) | The Ethics Committee Regione Toscana Area Great South East (CEAVSE), 20/04/2015, ref: HTRA1_GDF6_2015 |
| Health condition(s) or problem(s) studied | Age-related macular degeneration (AMD) |
| Intervention | We measure the concentrations of active TGF-β1, TGF-β2, and TGF-β3 by ELISA in the aqueous humor of patients affected by nAMD, who received 3 consecutive monthly intravitreal injections of ranibizumab according to the routine clinical practice management. Age-matched cataract patients serve as controls. Anterior chamber taps are performed in the operating room prior to each intravitreal injection (patients) and before cataract surgery (controls). A 30-gauge needle is inserted into the anterior chamber and 0.16-0.2 mL of aqueous is collected, centrifuged at 1500 g for 20 minutes to remove cells and debris, aliquoted and frozen at -80°C until analysis. Samples are collected at baseline (before the first injection), before the second injection, and before the third injection. The same samples are used in a luciferase-based reporter assay to test the TGF-β pathway activation. To perform the latter, Lenti-X 293T cells are transfected with a plasmid carrying the NanoLuc® gene under the transcriptional control of three copies of the SMAD binding element. The plasmid pGL4.54[luc2/TK] (Promega Corp., Madison, WI), carrying the reporter luc2 gene coding firefly luciferase under the control of the costitutive HSV-TK promoter, is co-transfected as a normalizer for transfection efficiency. Cells are then treated with 10 𝜇l of aqueous humor of the nAMD patients, naïve or treated, or of the control samples. After 3 hours Firefly and NanoLuc luciferase activities are measured for each sample. |
| Intervention type | Drug |
| Pharmaceutical study type(s) | |
| Phase | Not Applicable |
| Drug / device / biological / vaccine name(s) | Ranibizumab |
| Primary outcome measure | 1. TGF-beta concentration is measured using ELISA kits for human TGF-β1, TGF-β2 (Quantikine ELISA kits #DB100B and #DB250, respectively; R&D Systems, Minneapolis, USA) and TGF-β3 (#SEB949Hu; Cloud-Clone Corp., Houston, TX) at baseline, 1 month and 2 months (Only baseline for controls) 2. TGF-β pathway activation is measured using the transfection of Lenti-X 293T cells and the Nano-Glo® Dual-Luciferase® Reporter Assay System (#N1610; Promega Corp., Madison, WI) at baseline, 1 month and 2 months (only baseline for controls) |
| Secondary outcome measures | There were no secondary outcomes of interest. |
| Overall study start date | 16/03/2015 |
| Completion date | 21/06/2017 |
Eligibility
| Participant type(s) | Patient |
|---|---|
| Age group | Senior |
| Sex | Both |
| Target number of participants | 20 cases and 20 controls |
| Key inclusion criteria | Case inclusion criteria: 1. Both males and females 2. Patients aged 50 and over 3. Able to provide written, informed consent to the study 4. Active neovascular AMD in study eye with evidence of leakage from CNV on fluorescein angiography. Control inclusion criteria: 1. Both males and females 2. Patients aged 50 and over, the age of the controls is chosen not to have statistical differences with the cases 3. Able to provide written, informed consent to the study 4. Patients in the waiting list for cataract surgery |
| Key exclusion criteria | CASE exclusion criteria: 1. Prior treatment for neovascular age-related macular degeneration 2. Age less than 50 years old 3. Inability to comply with the study or follow up 4. Inability to perform fluorescein angiography 5. Axial length greater than 26 mm 6. Any previous ophthalmic surgery except cataract removal ( cataract surgery had to have been performed at least 9 months prior to inclusion) 7. Any other ocular disease other than neovascular age-related macular degeneration 8. Diabetes mellitus, use of immunosuppressive drugs and a malignant tumor in any location Control exclusion criteria: 1. Any other ocular disease other than cataract 2. Any previous ophthalmic surgery 3. Age less than 50 years old 4. Axial length greater than 26 mm 5. Diabetes mellitus, use of immunosuppressive drugs and a malignant tumor in any location |
| Date of first enrolment | 01/06/2016 |
| Date of final enrolment | 21/12/2016 |
Locations
Countries of recruitment
- Italy
Study participating centre
Surgery and Neuroscience
Viale Bracci, 1
Siena
53100
Italy
Sponsor information
University/education
Via Banchi di Sotto, 55
Siena
53100
Italy
| Phone | +39 0577 235555 |
|---|---|
| urp@unisi.it | |
| Website | https://en.unisi.it |
"ROR" |
https://ror.org/01tevnk56 |
Funders
Funder type
University/education
No information available
Results and Publications
| Intention to publish date | 01/06/2018 |
|---|---|
| Individual participant data (IPD) Intention to share | Yes |
| IPD sharing plan summary | Available on request |
| Publication and dissemination plan | Planned publication in a high-impact peer reviewed journal. |
| IPD sharing plan | The datasets generated during and/or analysed during the current study are/will be available upon request from Dr. Galvagni Federico Department of Biotechnology Chemistry and Pharmacy University of Siena via Aldo Moro, 53100 Siena, Italy e-mail: federico.galvagni@unisi.it |
Study outputs
| Output type | Details | Date created | Date added | Peer reviewed? | Patient-facing? |
|---|---|---|---|---|---|
| Results article | results | 23/05/2018 | Yes | No |
Editorial Notes
10/12/2018: Publication reference added.
