CNR

A diagnostic marker for Paget disease

The present invention relates to a new diagnostic marker for Paget’s disease of bone (PDB) and bone tumors associated therewith, such as Giant Cell Tumor (GCT). This marker is represented by the ZNF687 gene that, when mutated, is responsible for an aggressive form of PDB. If not treated, these PDB patients usually undergo the neoplastic degeneration.

Patent status

SUBMITTED

Priority Number

TO2014A000404

Priority Date

21/05/2014

License

INTERNATIONAL

Market

Paget disease of bone (PDB) is the second most common bone remodeling disease after osteoporosis. It is a chronic and disabling disease, affecting up to 1% of middle-aged patients (~50 years old). PDB prevalence notably differs amongst different Countries. This disease is most common (5,4%) in the United Kingdom, less in Japan (0,00028%).

Problem

Paget's disease of bone is a metabolic disorder that affects the skeletal system and is due to a dysregulation between the two types of cells that contribute to bone metabolism: osteoclasts (responsible for bone resorption) and osteoblasts (responsible for bone formation). This unbalance results in pain, deformity and fractures. However, the most fearful complication is represented by the tumor degeneration of the affected bones into osteosarcoma or, more rarely, giant cell tumor. A pharmacological treatment is available, using next generation bisphosphonates, which are able to block the evolution of the disease, especially if detected early. It is therefore absolutely important to diagnose the disease on time.
Recently, we found that patients with the very aggressive form of the disease that degenerates into giant cell tumor are mutated in the ZNF687 gene. The development of a diagnostic kit would make it possible to identify patients carrying the mutation early and subject them to prompt drug treatment, thus avoiding the onset of giant cell tumor.

Current Technology Limitations

Paget's disease is often asymptomatic in early stages and symptoms generally arise in the late stage: the most common symptom is bone pain, which can occur in one or more skeletal areas. Bone deformities and fractures typically arise in the advanced stage of the disease. For diagnostic confirmation, instrumental investigations such as radiography, bone scintigraphy and the dosage of some markers of bone metabolism are necessary (the most common is total alkaline phosphatase), generally increased in case of disease.
All these clinical and instrumental analyses can be anticipated by the molecular analysis that identifies carriers of the mutation in their early stage when a pharmacological treatment can block and/or control its pathological course.
The ZNF687 gene, responsible for a severe form of Paget’s disease of bone complicated by giant cell tumor degeneration, has been identified in the Molecular Genetics and Genomics laboratory at CNR of Naples in 2016. Previously, there was no diagnostic markers for this disorder.

Killer Application

The diagnostic procedure is based on the combination of different molecular assays: DNA extraction, amplification of the gene of interest by polymerase chain reaction (PCR), nucleotide sequencing by Sanger method.
This workflow would lead to the development of a molecular tool that, with few steps and few reagents, may be able to make an accurate molecular diagnosis of the disease.

Our Technology and solutions

We propose the development of a diagnostic kit able to detect the presence of mutations in the ZNF687 gene in patients affected by Paget's disease of the bone.
Through a blood sample, it will be possible to extract the DNA from the patient, subject to molecular analyses and finally sequence the causative gene.

The genetic test consists in the sequencing of the entire ZNF687, with particular attention to the exon 6 of the same gene, where the P937R mutation that causes giant cell tumor degeneration may be present.

Advantages

The main advantage is the possibility of identifying patients with Paget's disease of bone with a mutation in the ZNF687 gene and therefore predisposed to the development of giant cell tumor. A prompt and adequate pharmacological treatment of these patients could avoid the onset of giant cell tumor.

Two categories of patients could benefit from the existence of a diagnostic kit:
1. relatives of patients affected by Paget's disease of bone and mutated in the ZNF687 gene to verify their potential carrier status which can therefore be monitored along the time.
2. pagetic patients with a clinical diagnosis who may be predisposed to tumor development.

Roadmap

Interaction with pharmaceutical companies interested and/or involved in bone metabolism disorders and/or in neoplastic disorders for the development of the diagnostic kit.

An idea for the development of the diagnostic kit could include several steps:
a tube could contain all the enzymes for DNA extraction.
a second tube could contain enzymes to allow gene PCR amplification.
a third tube containing enzymes for DNA sequencing.

Development stage

The European patent n. 15732388 was deposited in Italy, France, Germany, United Kingdom and Belgium. It represents the starting point for an interaction with pharmaceutical companies interested in bone metabolism disorders and/or neoplastic disorders for the development of a diagnostic kit for the identification of mutations in the ZNF687 gene. This could be used by public or private hospitals and/or genetic diagnosis Centers.

There are currently no ongoing negotiations with companies for the development of an experimental proof of concept for the early diagnosis of this form of Paget's disease associated with neoplastic transformation in giant cell tumor.

The Technology Readiness Level can be considered at a level of TRL 2 (formulation of a technological concept).

Review the Technology

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Team

Fernando Gianfrancesco

He is a Senior Researcher at the Institute of Genetics and Biophysics at the National Research Council of Italy (IGB-CNR) in Naples. He is also Professor of Molecular Biotechnology at University of Campania Luigi Vanvitelli. He carried out research activities in several Research Centers of foreign Countries: USA, Australia, Portugal and United Kingdom. The scientific activity of Dr. Gianfrancesco has resulted in the publication of more than 80 original articles and book chapters. He is a member of several editorial boards as well as a regular reviewer of multiple scientific journals. Fernando Gianfrancesco’s scientific activity is focused on the study of human skeletal disorders and bone tumors. Besides the ZNF687 gene, responsible for a severe form of Paget’s disease of bone associated with giant cell tumor, his Lab recently identified the PFN1 gene, responsible for Osteosarcoma degeneration, sometimes linked to PDB.

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Federica Scotto di Carlo

She is a junior post-doc in the Lab of Dr. Gianfrancesco. She finished her PhD in 2018, during which she identified the loss of Profilin 1 as responsible for osteosarcoma transformation in patients with Paget’s disease. Thanks to this discovery, she was rewarded with several prizes, including the “Raisz-Drezner First Paper Award” and the “Felix Bronner Young Investigator Award” awarded by the American Society for Bone and Mineral Research as well as the “New Investigator Award” awarded by the European Calcified Tissue Society. She is currently focusing her attention on understanding how errors during DNA replication occur and cause cancer, especially osteosarcoma.

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Sharon Russo

She graduated in 2019 and she is at her first year of PhD course at the Institute of Genetics and Biophysics of the CNR in Naples. She is well experienced in mouse phenotypic characterization, and her main field of interest is the analysis of genetically mutated mice with the purpose of identifying phenotypic abnormalities, especially associated with bone disorders and tumors.

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