nir-vana innovation network

The University from Northern Italy is one of the largest Italian Universities, with 27 departments and over 75.000 students. It carries out scientific research and organizes courses in all disciplines, except for Engineering and Architecture. Biotechnology and medical sciences are only some of the areas in which the University excels.

Mutations in the "Isocitrate dehydrogenase 2" (IDH2) gene have been observed in several pathologies including acute myeloid leukemia (AML) and myelodysplasia (MDS). The two most frequent IDH2 mutations in AML and MDS affect more than 95% of mutated patients.
Also, the persistence of IDH2 mutations was observed in 40% of patients with AML in complete remission and is associated with an increased risk of recurrence. This suggested the role of IDH2 mutations as molecular markers of minimal residual disease, especially in the absence of other alterations. In addition, a specific drug for IDH2 mutated AML patients has recently been approved. For these reasons, it is essential to monitor the status of IDH2 to better characterize patients and to early direct them to the best therapy. Currently, to evaluate the status of IDH2 in these patients, Sanger sequencing is the most used method, but it is negatively characterized by its high detection limit (~ 20%) and by the need for expensive equipment and reagents.
For these reasons, some researchers of the University developed a diagnostic kit, based on a PNA-PCR Clamping (Peptide Nucleic Acid - Polymerase Chain Reaction Clamping), able to identify the 2 most frequent IDH2 mutations quickly, accurately and economically. This kit could allow the molecular characterization of IDH2 even in laboratories that currently cannot do it.

The kit was initially developed for the diagnosis and prognostic evaluation of patients with AML and MDS, but it can also be used to evaluate the mutational status of the IDH2 gene in any other pathology in which these mutations are present (e.g.: glioma, adenocarcinoma of the stomach, adenocarcinoma of the biliary tract, sarcoma, etc.).

The University is looking for start-ups and companies that can give technical and logistic support in the realization and commercialization of the kit.
Also, they are seeking Legal advice, to apply to European IVD certification, and Marketing expert for product promotion.

Advantages & innovations

The PNA-PCR Clamping method is an efficient alternative to conventional technics in the evaluation of mutations of the IDH2 gene. The kit allows the identification of the R140 and R172 mutations of the IDH2 gene using two simple PCR reactions. Compared to Sanger sequencing, which is currently the most used method, this kit: • is more sensitive and accurate (sensitivity: 87.5% vs. 66.7% and accuracy: 96.9% vs. 92.3%, respectively for PNA-PCR Clamping and Sanger sequencing); • has a lower limit of detection (1% vs. 20%, respectively for PNA-PCR Clamping and Sanger sequencing), which allows detecting even small percentages of mutated cells; • is faster (estimated time for PNA-PCR Clamping is 3 hours vs. Sanger sequencing 7 hours (excluding DNA extraction, common the other conventional techniques); • is cheaper (estimated reagent cost for analyzed DNA using PNA-PCR Clamping is €3 vs. Sanger sequencing €38 [excluding DNA extraction and electrophoretic running, common to the other conventional techniques] and estimated cost for equipment using PNA-PCR Clamping is €6.000 vs. Sanger sequencing €250.000); • high number of samples that can be analyzed at the same time; • simplicity of data interpretation ( does not require highly specialized personnel); • within the reach of all diagnostic laboratories, since it requires simple and inexpensive tools (thermocycler and apparatus for electrophoretic running, also necessary for Sanger sequencing).

Stage of development

Under development/lab tested

Partner sought

Partners needed (companies or startups): • Technical and logistic support to produce the kit; • Legal advice, to obtain the European IVD certification; • Marketing expert for product promotion.