Establishment of Genomic, Transcriptomic and Functional Characteristics of Tumor Cells in Hyperinflammatory Hemopathies
Establishment of Genomic, Transcriptomic and Functional Characteristics of Tumor Cells in Hyperinflammatory Hemopathies
Patients with monoclonal gammopathies either benign or malignant may develop inflammatory
 symptoms such as Schnitzler syndrome. We proposed to name those cases as monoclonal
 gammopathy of inflammatory significance, or MGIS.

 The aim of the study is to search for inflammatory pathway that may be activated in case MGIS
 and to search if the genetic of tumoral cell could explain those activations.
Auto-inflammatory Diseases (AIDs) are defined by disorders of the innate immunity. They
 manifest themselves as episodes of inflammation associated with general signs and organ
 damage, mainly to the cutaneous, musculoskeletal and digestive systems. The majority of
 monogenic AIDs described to date begin in childhood. Aside from impaired quality of life, and
 disease-related organ damage (e.g. deafness in periodic cryopyrin-associated syndromes), the
 most severe complication of AIDs is inflammatory amyloidosis (AA), which can lead to kidney
 failure and dialysis. More recently, late-onset AIDs have been described, secondary to
 somatic mutations in auto-inflammatory genes, mainly NLRP3 (1). This discovery comes in a
 more general context of highlighting new forms of late-onset genetic diseases linked to
 somatic mutations. These discoveries have been made possible by the revolution in sequencing
 techniques and in particular by the new generation sequencing which allows a more in-depth
 analysis compared to Sanger's technique.

 Blood neoplasms, known as clonal hemopathies, whether myeloid (such as myelodysplastic
 syndrome) or lymphoid (such as multiple myeloma), are constantly increasing in the general
 population. They are associated with an accumulation of somatic gene mutations that confer
 survival and proliferation properties to tumor cells (2). A number of manifestations of these
 clonal hemopathies are related to the activation of inflammatory pathways and in particular
 the activation of an inflammasome. For example, it has been shown that pyroptosis via
 activation of the NLRP3 inflammasome could be one of the pathophysiological mechanisms of
 myelodysplastic syndromes (3). In addition, inflammatory cytokines such as IL1 and IL6 have
 been shown to play a role as growth factors in these hemopathies (4). However, the mechanisms
 behind the activation of the inflammasome and the production of those cytokines are currently
 unknown.

 A number of patients with clonal hemopathies develop symptoms and complications similar to
 those encountered during AIDs. In particular, within the reference center for
 autoinflammatory diseases and amyloidosis in adults, we have identified patients with
 myelodysplastic syndromes or clonal gammopathies complicated by autoinflammatory symptoms or
 AA amyloidosis. These manifestations in the context of clonal hemopathies could be called
 "hyperinflammatory hemopathies ".

 Our hypothesis is that the inflammatory manifestations occurring in these patients are
 secondary to the occurrence of somatic mutations in genes known to be involved in innate
 immunity and AIDs.

 Our main objective is to perform in patients with hyperinflammatory hemopathy a functional
 study in search of the activation of inflammatory pathways; then a parallel genomic and
 transcriptomic study using an innovative methodology (5), in order to investigate whether
 there are somatic mutations of innate immunity genes in a clonal subpopulation and their
 impact on gene expression.

 The first step will be to identify the inflammatory signature of mononuclear blood cells from
 patients. To do this, we will recover the cells from blood and bone marrow. Real-time
 quantitative ELISA (Enzyme Linked ImmunoSorbent Assay) and PCR (Polymerase Chain Reaction)
 experiments will quantify the production of pro-inflammatory cytokines. In addition, flow
 cytometry experiments with membrane and intracellular co-labelling will be performed. These
 experiments, performed routinely in the laboratory, will make it possible to identify the
 cell type responsible for the overproduction of cytokines that may explain the symptoms.

 The second step, truly innovative and involving state-of-the-art techniques, will consist in
 carrying out genomic and transcriptomic analysis of tumor cells. For this we will use methods
 known as "single cell analysis" with parallel sequencing of the genome and transcriptome. The
 aim of genomic analysis is to search for somatic mutations present in tumor cells, and to
 compare them with those known to be associated with AIDs (comparison with the European
 Infevers database in particular. The genomic results will also be compared with international
 cancer cell databases to look for mutations in tumor cells that could explain this clinical
 presentation. These Big Data studies are necessary in view of the high number of mutations
 expected in tumor cells. The association with the parallel transcriptomic study will allow an
 unsupervised clustering analysis to study the effect of somatic mutations on gene expression.
 Indeed, it is currently recognized that the consequences of a genetic mutation must be
 analyzed both by its effect on the protein structure, but also by its effect on epigenetics.
 Today, this type of analysis is possible using the artificial intelligence analysis method.
Inclusion Criteria:

 -  Patient over 18 years of age with the capacity to give express free and informed
 consent.

 -  Patient followed in one of the services participating in the study and meeting the 2
 following conditions

 1. have myelodysplastic syndrome, or multiple myeloma, or monoclonal gammopathy of
 undetermined significance, or Schnitzler's syndrome.

 2. have a fever of undetermined etiology associated with one of the following
 symptoms: rash, arthralgia or arthritis, serositis, abdominal pain.

 Exclusion Criteria:

 -  Patient incapable of giving express free and informed consent.

 -  Subject under guardianship, curatorship or safeguarding of justice.

 -  Subject does not speak French.

 -  Subject incapable of answering questions or expressing himself/herself.

 -  Presence of a differential diagnosis that may explain the patient's symptoms.