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US-12617835-B2 - Chimeric autoantibody receptor (CAAR) that binds autoantibodies targeting the central nervous system in neurological autoimmune disease

US12617835B2US 12617835 B2US12617835 B2US 12617835B2US-12617835-B2

Abstract

A chimeric autoantibody receptor (CAAR) that enables targeting of an immune cell to autoantibody producing B cells. The CAAR includes an autoantigen or fragment thereof that is bound by autoantibodies associated with neurological autoimmune disease primarily targeting the central nervous system. Also disclosed is a nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR), the nucleic acid sequence encoding an autoantigen or fragment thereof that is bound by autoantibodies associated with a neurological autoimmune disease primarily targeting the central nervous system, a transmembrane domain, and an intracellular signaling domain, a vector comprising a nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR), a genetically modified immune cell comprising the nucleic acid molecule encoding the CAAR and use of the immune cell in the treatment or prevention of a neurological autoimmune disease primarily targeting the central nervous system, such as an autoimmune encephalopathy or encephalomyelopathy, preferably anti-NMDAR encephalitis.

Inventors

  • Harald Prüss
  • S. Momsen Reincke
  • Inan Edes

Assignees

  • DEUTSCHES ZENTRUM FÜR NEURODEGENERATIVE ERKRANKUNGEN E. V. (DZNE)
  • MAX-DELBRÜCK-CENTRUM FÜR MOLEKULARE MEDIZIN IN DER HELMHOLTZ-GEMEINSCHAFT

Dates

Publication Date
20260505
Application Date
20200605
Priority Date
20190605

Claims (11)

  1. 1 . A nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR), the nucleic acid molecule comprising: i. a sequence encoding an autoantigen comprising an amino-terminal domain (ATD) of an NMDA receptor according to SEQ ID NO 14, or a fragment thereof that is bound by autoantibodies in anti-N-methyl-D-aspartate receptor encephalitis (anti-NMDAR encephalitis), ii. a sequence encoding a transmembrane domain, and iii. a sequence encoding an intracellular signaling domain.
  2. 2 . The nucleic acid molecule according to claim 1 : wherein the transmembrane domain is a CD8 alpha transmembrane domain; wherein the intracellular domain comprises a CD137 (4-1BB) co-stimulatory domain; wherein the intracellular domain comprises a CD3 zeta chain signaling domain; and wherein the nucleic acid molecule comprises additionally one or more sequences encoding one or more leader, linker and/or spacer polypeptides positioned between the autoantigen and transmembrane domain and/or N-terminally of and/or between fragments of the autoantigen, and/or between the transmembrane and intracellular co-stimulatory domain.
  3. 3 . The nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR) according to claim 1 , comprising: i. a sequence encoding a leader polypeptide, wherein the leader polypeptide is a CD8 leader polypeptide; ii. a sequence encoding an autoantigen according to claim 1 ; iii. optionally a sequence encoding a linker polypeptide positioned between one or more NMDAR fragments; iv. optionally a sequence encoding a linker polypeptide positioned between the autoantigen and transmembrane domain; V. a sequence encoding a CD8 alpha transmembrane domain; vi. optionally a sequence encoding a linker polypeptide positioned between a transmembrane domain and an intracellular signaling domain; and vii. a sequence encoding an intracellular signaling domain, said intracellular signaling domain comprising a CD137 (4-1BB) co-stimulatory domain and a CD3 zeta chain signaling domain, wherein optionally a linker sequence is positioned between the co-stimulatory and signaling domains.
  4. 4 . The nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR) according to claim 1 , further comprising one or more of: a sequence encoding a leader polypeptide, wherein the leader polypeptide is a CD8 leader polypeptide, said sequence comprising a sequence according to SEQ ID NO 1; a sequence encoding an autoantigen according to claim 1 , said sequence comprising a sequence according to SEQ ID NO 3 (ATD); a sequence encoding a linker polypeptide positioned between one or more NMDAR fragments, said sequence comprising a sequence according to GGCACC (linker-1); a sequence encoding a linker polypeptide positioned between the autoantigen and transmembrane domain, said sequence comprising a sequence according to SEQ ID NO 7 (linker-2) or SEQ ID NO 32 (linker-2b); a sequence encoding a CD8 alpha transmembrane domain according to SEQ ID NO 8; a sequence encoding a linker polypeptide positioned between a transmembrane domain and an intracellular signaling domain, said sequence comprising a sequence according to GGCAGC (linker-3); and a sequence encoding an intracellular signaling domain, said intracellular signaling domain comprising a CD137 (4-1BB) co-stimulatory domain and a CD3 zeta chain signaling domain, said sequence comprising a sequence according to SEQ ID NO 10 (CD137) and SEQ ID NO 11 (CD3z), respectively, wherein optionally a linker sequence is positioned between the co-stimulatory and signaling domains.
  5. 5 . The nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR) according to claim 1 , wherein the autoantigen encoded by the nucleic acid sequence comprises one or more fragments of a NR1 and/or NR2 subunit of a NMDA receptor and not a complete NR1 and/or NR2 subunit.
  6. 6 . The nucleic acid molecule according to claim 1 , wherein the autoantigen comprises an amino-terminal domain (ATD) of an NMDA receptor according to SEQ ID NO 14.
  7. 7 . The nucleic acid molecule according to claim 6 , wherein the transmembrane domain is a CD8 alpha transmembrane domain; and wherein the intracellular domain comprises a CD137 (4-1BB) co-stimulatory domain and a CD3 zeta chain signaling domain.
  8. 8 . The nucleic acid molecule according to claim 7 , wherein the transmembrane domain is a CD8 alpha transmembrane domain according to SEQ ID NO 20; and wherein the intracellular domain comprises a CD137 (4-1BB) co-stimulatory domain according to SEQ ID NO 22 and a CD3 zeta chain signaling domain according to SEQ ID NO 23.
  9. 9 . The nucleic acid molecule according to claim 1 , wherein the transmembrane domain and the intracellular domain comprise an ICOS transmembrane domain and ICOS intracellular domain according to SEQ ID NO 21.
  10. 10 . A vector comprising a nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR) according to claim 1 .
  11. 11 . A vector according to claim 10 , wherein the vector is a viral vector, nanoparticles as a transfection vehicle, a transposon or an RNA vector.

Description

The invention relates to the field of targeted cellular therapy employing a chimeric autoantibody receptor and the treatment of neurological autoimmune disease. The invention relates to a chimeric autoantibody receptor (CAAR) that enables targeting of an immune cell to autoantibody producing B cells. The CAAR comprises an autoantigen or fragment thereof that is bound by autoantibodies associated with a neurological autoimmune disease primarily targeting the central nervous system. The invention relates to a nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR), the nucleic acid molecule comprising a sequence encoding an autoantigen or fragment thereof that is bound by autoantibodies associated with a neurological autoimmune disease primarily targeting the central nervous system, a sequence encoding a transmembrane domain, and a sequence encoding an intracellular signaling domain. In one embodiment, the autoantigen encoded by the nucleic acid sequence comprises or consists of an N-methyl-D-aspartate receptor (NMDAR), or one or more NMDAR fragments. The invention further relates to the chimeric autoantibody receptor (CAAR) protein of the invention, a vector comprising a nucleic acid molecule encoding a chimeric autoantibody receptor (CAAR) of the invention, a genetically modified immune cell comprising the nucleic acid molecule encoding the CAAR and the use of the immune cell in the treatment or prevention of a neurological autoimmune disease primarily targeting the central nervous system, such as an autoimmune encephalopathy or encephalomyelopathy, preferably anti-NMDAR encephalitis. REFERENCE TO SEQUENCE LISTING The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 54687469_1.TXT, created and last modified on Dec. 3, 2021, which is 61.7 Kb in size. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety. BACKGROUND OF THE INVENTION Autoimmunity, a main component in nervous system disease, is a misguided immune response to the body's own organs. Neurological autoimmune disease occurs when autoimmunity (autoantibodies) targets a structure within the central or peripheral nervous system. Anti-N-methyl-D-aspartate receptor encephalitis (anti-NMDAR encephalitis) has recently been discovered as an autoimmune neuropsychiatric disease in which autoantibodies against the NR1 subunit of the NMDA receptor are formed and bind to NMDA receptors (NMDAR) in the brain (Titulaer 2013). Binding of the autoantibodies to the NMDAR leads to an internalization of the receptors and thus to a dysfunction of affected nerve cells (Kreye 2016), commonly characterized by symptoms such as epileptic seizures, impaired consciousness, movement disorders, memory loss and signs of psychosis (Dalmau 2011, Prüss 2017). The removal of the autoantibodies from the blood and cerebrospinal fluid of patients leads to a significant clinical improvement, such that many patients can lead an independent life after appropriate treatment removing said autoantibodies. However, significant problems are associated with established therapeutic approaches using unspecific immunosuppression, such as steroid treatment, plasmapheresis, cyclophosphamide or rituximab treatment (to deplete antibody-producing B-cells). These treatments have led to an improvement in the patient's condition but are associated with significant side effects (Titulaer 2013). In the case of plasmapheresis, unwanted side effects typically arise as injuries caused by the central venous catheter, the development of circulatory disorders such as hypotensive and/or circulatory dysregulation, i.e. due to fluid shifting, coagulation disorders with thrombosis, and infections, including sepsis. Drug-induced immunosuppression is especially susceptible to causing severe infections, in addition to the sometimes significant well-known side effects of pharmacological therapy. Furthermore, prophylaxis by vaccinations and the protection of the body by beneficial antibodies that challenge bacterial and viral infection can be negated by unspecific immunotherapies. For example, the removal of autoantibodies in general does not lead per se to the removal of the autoantibody-producing cells, i.e. the source of the disease-causing agent. In the acute phase of anti-NMDAR encephalitis, substantial quantities of disease-causing autoantibodies are produced by the responsible B cells. This production is not inhibited by removal of the autoantibodies, as long as the responsible B cells remain active, thereby leading to the necessity of repeated autoantibody-removal procedures, such as apheresis and the like. These problems can only be solved by selective approaches for removal of disease-specific autoantibodies and the cause of their production. Until the present time, to the best knowledge of the inventors, no effective therapeutic options are availab