Detection of hepatitis delta virus (HDV) for the diagnosis and treatment of Sjögren's syndrome and lymphoma
Inventors
Weller, Melodie L. • Chiorini, John A.
Assignees
US Department of Health and Human Services
Publication Number
US-10584387-B2
Publication Date
2020-03-10
Expiration Date
2034-10-09
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Viral infection has been suspected in the development of primary Sjögren's syndrome (pSS). Using a custom viral microarray, hepatitis delta virus (HDV) genomes and antigens were detected in minor salivary glands of patients with pSS. Expression of HDV antigens in healthy mice led to a Sjögren's syndrome-like pathogenesis characterized by a reduction in salivary flow, increase in lymphocytic foci, and the development of an autoantibody profile similar to HDV-positive patients. Also described herein is the detection of HDV in patients diagnosed with lymphoma. Expression of HDV antigen in healthy mice resulted in the development of tertiary lymphoid structures characteristic of the early stages of lymphoma. A sensitive, nested qPCR assay to detect HDV transcript and/or HDV genome in patient samples is also described.
Core Innovation
The invention identifies hepatitis delta virus (HDV) nucleic acid and antigens in the salivary glands of patients diagnosed with primary Sjögren's syndrome (pSS) and in lymphoma tumor biopsies. It demonstrates that expression of HDV antigens in healthy mice induces a Sjögren's syndrome-like pathogenesis characterized by reduction in salivary flow, increased lymphocytic foci, and autoantibody profiles similar to HDV-positive patients. Expression of HDV antigen in healthy mice also results in development of organized tertiary lymphoid structures indicative of early-stage lymphoma.
The invention provides methods of diagnosing a subject as having or being susceptible to Sjögren's syndrome or lymphoma by detecting HDV nucleic acid or antigen in a sample obtained from the subject. It further provides treatment methods that include administering an inhibitor of HDV to subjects diagnosed with or susceptible to these diseases. The invention also describes isolated nucleic acid molecules, oligonucleotides, primers, probes specific for HDV detection, and kits for detecting HDV nucleic acid in biological samples.
The problem being solved addresses the unmet need to identify viral triggers linked to the development of primary Sjögren's syndrome and lymphoma. Prior studies suggested viral involvement but failed to establish a direct association between viral infection and autoimmune disease development. There was a lack of sensitive and specific assays to detect HDV in relevant patient tissues. Moreover, the etiology of lymphoma in pSS patients remained unidentified. The invention resolves these issues by providing sensitive detection methods and demonstrating a causative role for HDV in pSS and lymphoma pathogenesis, enabling novel diagnostic and therapeutic approaches.
Claims Coverage
The patent presents three independent claims each covering distinct inventive features related to diagnosing and treating Sjögren's syndrome and non-Hodgkin's lymphoma through detection of HDV nucleic acid or antigen.
Method of diagnosing Sjögren's syndrome by detecting HDV nucleic acid and autoantibodies or lymphocytic infiltrates
A method comprising detecting HDV nucleic acid in a salivary gland sample by RT-PCR using HDV-specific primers (including SEQ ID NOs: 3, 4, 11-14), detecting auto-antibodies to Ro(SSA) and/or La(SSB) in blood/serum or detecting focal lymphocytic infiltrates in the salivary gland, and diagnosing Sjögren's syndrome if both HDV nucleic acid and either autoantibodies or lymphocytic infiltrates are detected.
Nested PCR assay for sensitive detection of HDV transcript
The RT-PCR assay comprises a nested PCR with a first round using a first primer pair (SEQ ID NOs: 11 and 12) and a second round using a second primer pair (SEQ ID NOs: 13 and 14), amplifying HDV transcript if present, and detecting the transcript with a probe comprising SEQ ID NO: 15.
Method of treating Sjögren's syndrome after diagnosis by detecting HDV nucleic acid and autoantibodies or lymphocytic infiltrates
A method including detecting HDV nucleic acid in a salivary gland sample, detecting autoantibodies or lymphocytic infiltrates, diagnosing Sjögren's syndrome, and administering therapy comprising agents that promote salivary production, corticosteroids, immunosuppressive drugs, NSAIDs, or combinations thereof.
Method of diagnosing and treating non-Hodgkin's lymphoma by detecting HDV nucleic acid and tertiary lymphoid structures
A method comprising detecting HDV nucleic acid in a salivary gland sample via RT-PCR using HDV-specific primers, detecting tertiary lymphoid structures in the sample, diagnosing non-Hodgkin's lymphoma if both are present, and administering appropriate therapy including radiation, chemotherapy, stem cell transplant, immunotherapy, surgery or combinations thereof.
Method of diagnosing susceptibility to non-Hodgkin's lymphoma by detecting HDV nucleic acid and tertiary lymphoid structures
A method comprising detecting HDV nucleic acid by RT-PCR using HDV-specific primers and detecting tertiary lymphoid structures in a salivary gland sample, and diagnosing susceptibility to non-Hodgkin's lymphoma if both are detected.
The independent claims cover diagnostic methods involving detection of HDV nucleic acid and viral antigens in patient samples, employing nested PCR assays with specific primers and probes. They include methods for diagnosing and treating Sjögren's syndrome and non-Hodgkin's lymphoma, based on presence of HDV and disease biomarkers. Treatment methods specify administration of therapies including agents to restore salivary function and conventional lymphoma treatments after HDV-based diagnosis.
Stated Advantages
Identification of HDV as a viral trigger enabling etiological diagnosis of Sjögren's syndrome and lymphoma.
Sensitive nested PCR assay provides improved specificity and sensitivity for HDV detection in patient samples.
Treatment methods targeting HDV infection offer new therapeutic avenues for Sjögren's syndrome and lymphoma.
Animal model demonstrates causative role of HDV antigen expression in disease phenotypes, supporting targeted intervention.
Documented Applications
Diagnosis of primary Sjögren's syndrome by detecting HDV nucleic acid or antigen in salivary gland tissue and related serological markers.
Treatment of Sjögren's syndrome by administration of HDV inhibitors or agents promoting salivary production.
Diagnosis of non-Hodgkin's lymphoma, including mucosa-associated lymphoid tissue (MALT) lymphoma, via detection of HDV nucleic acid and tertiary lymphoid structures.
Treatment of lymphoma by administration of HDV inhibitors and standard oncological therapies such as chemotherapy, radiation, immunotherapy, and surgery.
Use of nested quantitative PCR assays and immunoassays for HDV detection in biological samples including salivary gland biopsies, saliva, blood, and tumor biopsies.
Interested in licensing this patent?