Effect of B-cell Depleting Therapies on PLA2R-specific B Cells in Patients With Membranous Nephropathy

NCT06994468 | Completed

• 1 other identifier: DEEPER
• Study Type: interventional
• Target: 48
• Locations: 1 country
• Sites: 1

Brief Summary

This is a non-pharmacological interventional monocentric study to primarily assess the levels of circulating phospholipase A2 receptor (PLA2R)-specific B cells in patients with PLA2R-mediated MN enrolled in the ORION and MONET studies, before and at the different time points after therapy. It will also compare the phenotype of circulating PLA2R-specific B cell subsets over time in patients who achieved long-term remission, who experience MN relapses or did not respond to Obinutuzumab or Felzartamab therapies, and evaluate changes in lymphocyte subpopulations, including T cells and NK cells, possibly involved in the autoimmune process and in the response to B-cell depleting therapy.

Conditions

Membranous Nephropathy

Keywords

primary membranous nephropathy; PLA2R-specific B cells; Obinutuzumab; Felzartamab

Outcome Measures

Primary Outcomes (2)

• Levels of circulating PLA2R-specific B cells

  Levels of circulating PLA2R-specific B cells in patients with PLA2R-mediated MN enrolled in the ORION study

  Levels measured at baseline and at 3, 6, 9, 12, 18, 24 months.

• Levels of circulating PLA2R-specific B cells

  Levels of circulating PLA2R-specific B cells in patients with PLA2R-mediated MN enrolled in the MONET studies

  Levels measured at baseline, 29, 141 days and at 6, 9, 12, 18, 24 months.

Study Arms (5)

PBMCs from ORION patients positive for anti-PLAR2

EXPERIMENTAL

Peripheral Blood Mononuclear Cells (PBMCs) samples identified among anti-PLAR2 positive patients enrolled in the ORION study, who provided consent to store their samples collected during the follow-up at the Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò", Ranica (BG).

Diagnostic Test: Flow-cytometry tetramer-based antigen-bait assay

PBMCs from MONET patients positive for anti-PLAR2

EXPERIMENTAL

Peripheral Blood Mononuclear Cells (PBMCs) samples identified among anti-PLAR2 positive patients enrolled in the MONET study, who provided consent to store their samples collected during the follow-up at the Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò", Ranica (BG).

Diagnostic Test: Flow-cytometry tetramer-based antigen-bait assay

PBMCs from ORION patients negative for anti-PLAR2

ACTIVE COMPARATOR

Peripheral Blood Mononuclear Cells (PBMCs) samples identified among anti-PLAR2 negative patients enrolled in the ORION study, who provided consent to store their samples collected during the follow-up at the Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò", Ranica (BG).

Diagnostic Test: Flow-cytometry tetramer-based antigen-bait assay

PBMCs from MONET patients negative for anti-PLAR2

ACTIVE COMPARATOR

Peripheral Blood Mononuclear Cells (PBMCs) samples identified among anti-PLAR2 negative patients enrolled in the MONET study, who provided consent to store their samples collected during the follow-up at the Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò", Ranica (BG).

Diagnostic Test: Flow-cytometry tetramer-based antigen-bait assay

PBMCs isolated from volunteers

NO INTERVENTION

PBMCs isolated from volunteer subjects without a history of renal diseases, autoimmune disorders, diabetes mellitus, or current allergies, and who have undergone a one-week washout from antibiotics, anti-inflammatory medications, and antihistamines.

Interventions

Flow-cytometry tetramer-based antigen-bait assay DIAGNOSTIC_TEST

For tetramer assembly, recombinant biotinylated PLA2R will be incubated with either streptavidin-PE (Agilent) or streptavidin-APC (Agilent) conjugates according to the NIH protocol (https://tetramer.yerkes.emory.edu/support/protocols#10). PBMCs will be thawed and resuspended in antibody cocktail medium containing 7.5ug APC- and PE- PLA2R tetramers. Multicolor flow cytometry will be used to identify B-cell subsets with standard technique and equipment (FACS FortessaX20, BD Biosciences and FlowJo software). PLA2R-specific B cells will be determined as Tetramer double (PE/APC) positive B cells that bind with proportional fluorescent intensities to each tetramer. Each sample will be incubated with only streptavidin-PE/APC conjugates to exclude B cells binding to the fluorochrome portion of the tetramers. T cell- and NK/NKT cell phenotyping will be assessed by FACS using ad-hoc antibody panels.

PBMCs from MONET patients negative for anti-PLAR2; PBMCs from MONET patients positive for anti-PLAR2; PBMCs from ORION patients negative for anti-PLAR2; PBMCs from ORION patients positive for anti-PLAR2

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Patients enrolled in the ORION and MONET studies who have who provided written informed consent to store their samples collected during the follow-up at the Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò", Ranica (BG).

You may not qualify if:

• No written informed consent to store their samples collected during the follow-up at the Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò", Ranica (BG).
• Volunteers
• Adult males and females.
• Written informed consent.
• History of renal diseases, autoimmune disorders, diabetes mellitus, current allergies.
• Subjects who have taken antibiotics, anti-inflammatory drugs, or antihistamines within the past 7 days.

Sponsors & Collaborators

• Mario Negri Institute for Pharmacological Research: lead

Study Sites (1)

Clinical Research Centre for Rare Diseases Aldo e Cele Daccò

Ranica, BG, 24020, Italy

Location

Related Publications (3)

• Ruggenenti P, Debiec H, Ruggiero B, Chianca A, Pelle T, Gaspari F, Suardi F, Gagliardini E, Orisio S, Benigni A, Ronco P, Remuzzi G. Anti-Phospholipase A2 Receptor Antibody Titer Predicts Post-Rituximab Outcome of Membranous Nephropathy. J Am Soc Nephrol. 2015 Oct;26(10):2545-58. doi: 10.1681/ASN.2014070640. Epub 2015 Mar 24.

  PMID: 25804280 BACKGROUND

• Beck LH Jr, Fervenza FC, Beck DM, Bonegio RG, Malik FA, Erickson SB, Cosio FG, Cattran DC, Salant DJ. Rituximab-induced depletion of anti-PLA2R autoantibodies predicts response in membranous nephropathy. J Am Soc Nephrol. 2011 Aug;22(8):1543-50. doi: 10.1681/ASN.2010111125. Epub 2011 Jul 22.

  PMID: 21784898 BACKGROUND

• Beck LH Jr, Bonegio RG, Lambeau G, Beck DM, Powell DW, Cummins TD, Klein JB, Salant DJ. M-type phospholipase A2 receptor as target antigen in idiopathic membranous nephropathy. N Engl J Med. 2009 Jul 2;361(1):11-21. doi: 10.1056/NEJMoa0810457.

  PMID: 19571279 BACKGROUND

MeSH Terms

Conditions

Glomerulonephritis, Membranous

Condition Hierarchy (Ancestors)

Glomerulonephritis; Nephritis; Kidney Diseases; Urologic Diseases; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Urogenital Diseases; Male Urogenital Diseases; Autoimmune Diseases; Immune System Diseases

Study Officials

• Giuseppe Remuzzi, M.D.

  Istituto Di Ricerche Farmacologiche Mario Negri

  STUDY DIRECTOR

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NON RANDOMIZED
• Masking: NONE
• Purpose: DIAGNOSTIC
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Model Details: PBMCs will be isolated form both anti-PLA2 positive and anti-PLAR2 negative patients enrolled in the ORION and MONET studies. The anti-PLAR2 negative patients will be sued as negative control patients. In addition, PBMCs will be isolated from volunteers. These PBMCs will serve to set a reference range for the planned analyses, ensuring the absence of confounding factors related to underlying health conditions or pharmacological interventions.

Study Record Dates

• First Submitted: May 20, 2025
• First Posted: May 29, 2025
• Study Start: September 22, 2025
• Primary Completion: December 2, 2025
• Study Completion: December 2, 2025
• Last Updated: March 20, 2026

Record last verified: 2026-03

Data Sharing

• IPD Sharing: Will not share

Locations

IT (1)