Functional Implications of Rare Gene Mutations in aHUS Open the Door to Personalized Therapy (aHUS-iPSC-EC)

Hemolytic Uremic Syndrome (HUS) is a rare disease characterized by rupture of red blood cells (hemolytic anemia), low platelet count (thrombocytopenia), and thrombotic occlusion of small vessels (thrombotic microangiopathy), with prevalent involvement of the kidneys.

SEU, in its typical form is caused by gastrointestinal infection with Escherichia coli.

The atypical form of SEU (aSEU), which is not caused by an Escherichia coli infection, is a very rare disease that may have a genetic origin; it affects both children and adults and may occur in a sporadic or familial form. Many studies have shown that about 60% of cases of atypical HUS are associated with genetic abnormalities of the complement system (particularly the so-called "alternative pathway"), which is a key part of the immune system for responding to infection. Complement consists of a series of proteins that, when activated, create a so-called "cascade," which leads to the elimination of the infectious agent, either directly or through other cells. Complement is finely regulated in such a way as to prevent damage to healthy cells in one's own body. Genetic defects in some of these complement regulatory proteins cause reduced protection of the endothelial surface (thus the vessel wall) against complement activation.

Recently, new mutations have been described in a gene unrelated to the complement pathway, the DKGE gene, which codes for the intracellular isoform of diacylglycerol kinase . In these patients, small renal vessel occlusion appears to occur as a result of altered endothelial cell proliferation and angiogenesis through mechanisms apparently unrelated to complement activation. However, to date these mechanisms are poorly studied. Throughout the entire project statistical methods will be applied to optimize the characterization of the abnormalities in phenotype and function of iPSC-EC derived from aHUS patients with either DGKE or MCP genetic abnormalities as compared with control iPSC-EC, including identifying potential drugs that could correct the abnormalities

Study Overview

• Status: Recruiting
• Conditions: Atypical Hemolytic Uremic Syndrome
• Intervention / Treatment: Other: Blood sampling and urine analysis

• Study Type: Interventional
• Enrollment (Estimated): 112
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Marina Noris, Dr.; Phone Number: +3903545351; Email: marina.noris@marionegri.it

Study Locations

• Italy
  • BG
    • Ranica, BG, Italy, 24020
      • Recruiting
      • Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò"
      • Contact: Erica Daina; Phone Number: 0039 035 45351; Email: erica.daina@marionegri.it

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult; Older Adult
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• Adults and children with aHUS defined by history of microangiopathic hemolytic anemia and thrombocytopenia (hematocrit (Ht) <30%, hemoglobin (Hb) <10 g/dL, LDH >500 IU/L, undetectable haptoglobin, fragmented erythrocytes in the peripheral blood smear with negative Coomb's test, and platelet count <150,000/microL), associated with acute renal failure.
• Written informed consent

Exclusion Criteria:

• TTP (ADAMTS13 activity <10%)
• STEC-HUS (presence of stx and eae genes or Shiga-toxin in the stools and/or serum antibodies against Shiga-toxin and/or STEC LPS).
• Disseminated intravascular coagulation (prolonged thromboplastin time and lower than normal fibrinogen levels).

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Screening
• Allocation: Non-Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: aHUS Patient; The study will include 110 patients consenting adult and pediatric patients with a diagnosis of atypical hemolytic uremic syndrome and carrying mutations in the MCP or DGKE genes. New patients will be selected through clinical and genetic screening of the inhabitants of a small island of South Italy (Linosa) with high incidence of patients affected by DGKE mutations and characterized by a high rate of endogamy	Other: Blood sampling and urine analysis; A blood sample of 10- 20 ml from pediatric patients, 30-50 ml from adult patients will be collected for each patient and healthy voluntarees
Other: Healthy volunteer; 2 healthy subjects will undergo urine analysis (multistick) and only subjects with normal parameters will be enrolled as controls.	Other: Blood sampling and urine analysis; A blood sample of 10- 20 ml from pediatric patients, 30-50 ml from adult patients will be collected for each patient and healthy voluntarees

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Time Frame
Generation and characterization of patient-specific and healthy donor iPSC	once during the study
Differentiation of iPSC into endothelial cells	once during the study
Characterizationof iPSC into endothelial cells	once during the study
Cell culture viability	once during the study

Collaborators and Investigators

• Sponsor: Mario Negri Institute for Pharmacological Research

Study record dates

Study Major Dates

• Study Start (Actual): May 3, 2023
• Primary Completion (Estimated): December 31, 2026
• Study Completion (Estimated): December 31, 2026

Study Registration Dates

• First Submitted: March 28, 2023
• First Submitted That Met QC Criteria: March 28, 2023
• First Posted (Actual): April 7, 2023

Study Record Updates

• Last Update Posted (Estimated): September 29, 2025
• Last Update Submitted That Met QC Criteria: September 23, 2025
• Last Verified: September 1, 2025

More Information

Terms related to this study

• Keywords: Endothelial cells; Eculizumab; Induced pluripotent stem cells; Alternative complement pathway; Atypical hemolytic uremic syndrome; Drug screening (in vitro)
• Additional Relevant MeSH Terms: Urogenital Diseases; Cytopenia; Male Urogenital Diseases; Kidney Diseases; Urologic Diseases; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Hematologic Diseases; Anemia, Hemolytic; Anemia; Blood Platelet Disorders; Thrombotic Microangiopathies; Thrombocytopenia; Uremia; Hemic and Lymphatic Diseases; Hemolytic-Uremic Syndrome; Atypical Hemolytic Uremic Syndrome; Investigative Techniques; Specimen Handling; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Punctures; Surgical Procedures, Operative; Diagnostic Techniques, Urological; Clinical Chemistry Tests; Blood Specimen Collection; Urinalysis
• Other Study ID Numbers: aHUS-iPSC-EC

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No