BCMA-directed CAR-T cell therapy for multiple myeloma

Abstract

Dose of T cells for use in a procedure for the treatment of multiple myeloma in a subject, wherein the T cells comprise a chimeric antigen receptor (CAR), comprising: (a) an extracellular antigen-binding domain capable of binding specifically to an epitope of the B-cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD.

Description

CROSS-REFERENCE TO RELATED REGISTRATIONS

[0001] The application claims priority over the preliminary US patent applications No. 63 / 497,185, filed on April 19, 2023, and No. 63 / 504,184, filed on May 24, 2023, and the international patent application No. PCT / US2023 / 031673, filed on August 31, 2023, the disclosures of which are incorporated herein in their entirety by reference. SEQUENCE PROTOCOL

[0002] The application includes a machine-readable sequence protocol, which was submitted with this application in XML file format, the entire content of which is incorporated herein by reference. The sequence protocol XML file submitted with this application is titled “14651-066-228_SEQ_LISTING.xml”, was created on August 24, 2023, and is 28,450 bytes in size. BACKGROUND

[0003] Multiple myeloma is an aggressive neoplasm of plasma cells. It is considered a B-cell neoplasm that proliferates uncontrollably in the bone marrow. Symptoms include one or more of the following: hypercalcemia, renal failure, anemia, bone lesions, bacterial infections, hyperviscosity, and amyloidosis. Despite the availability of new therapies, including proteasome inhibitors, immunomodulatory drugs, and monoclonal antibodies, which have shown significantly improved patient outcomes, multiple myeloma remains a nearly incurable disease. Because most patients either relapse or become refractory to treatment, the need for new therapies for multiple myeloma persists.In particular, patients with multiple myeloma who have previously received 1-3 lines of therapy and are refractory to lenalidomide have a poor prognosis and rapidly progress with available therapies, highlighting the need for new, safe and effective treatment regimens for use in these earlier line settings. SUMMARY OF THE REVELATION

[0004] In one aspect, a method for treating a subject is provided, comprising administering a dose of T cells comprising a chimeric antigen receptor (CAR) to the subject, comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of the B-cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain. In some embodiments, the subject has multiple myeloma, has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD. In some embodiments, the subject has a high-risk feature, and optionally, the high-risk feature is a cytogenetic abnormality, International Staging System (ISS) stage III, and / or soft tissue plasmacytomas.

[0005] In one aspect, a method for the selective treatment of a subject is provided here, comprising: (1) determining whether the subject has a high-risk feature, wherein the high-risk feature is a cytogenetic abnormality, International Staging System (ISS) stage III, and / or soft tissue plasmacytomas; and (2) administering to the subject in whom the high-risk feature was identified in step (1) a dose of T cells comprising a chimeric antigen receptor (CAR) comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of BCMA, (b) a transmembrane domain, and (c) an intracellular signaling domain. In some embodiments, the subject has multiple myeloma, has received one to three prior lines of therapy, including therapy with an IMiD, and is refractory to the IMiD.

[0006] In one aspect, a method for the selective treatment of a subject is provided, comprising administering a dose of T cells comprising a chimeric antigen receptor (CAR) to the subject identified as having a high-risk feature comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of BCMA, (b) a transmembrane domain, and (c) an intracellular signaling domain. In some embodiments, the high-risk feature is a cytogenetic abnormality, International Staging System (ISS) stage III, and / or soft tissue plasmacytomas. In some embodiments, the subject has multiple myeloma, has received one to three prior lines of therapy, including therapy with an IMiD, and is refractory to the IMiD.

[0007] In some embodiments of the various methods or aspects provided here, the IMiD is lenalidomide.

[0008] In some embodiments of the various methods or aspects provided herein, the high-risk feature is a cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality. In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from a group that includes Gain / AMP(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic abnormality includes Gain / AMP(1q). In some embodiments, the cytogenetic abnormality includes del(17p). In some embodiments, the cytogenetic abnormality includes t(4;14). In some embodiments, the cytogenetic abnormality includes t(14;16). In some embodiments, the subject has at least two cytogenetic abnormalities. In some embodiments, the subject exhibits at least three cytogenetic abnormalities.In some embodiments, the subject has at least four cytogenetic abnormalities. In some embodiments, the subject has at least five cytogenetic abnormalities. In some embodiments, the subject has at least six cytogenetic abnormalities. In some embodiments, the subject has at least seven cytogenetic abnormalities. In other embodiments, the cytogenetic abnormality is a standard-risk cytogenetic abnormality. In other embodiments, the high-risk feature is International Staging System (ISS) Level III. In still other embodiments, the high-risk feature is soft tissue plasmacytomas.

[0009] In some embodiments of the various methods or aspects provided here, the subject has received one prior line of therapy. In some embodiments, the subject has received two prior lines of therapy. In some embodiments, the subject has received three prior lines of therapy.

[0010] In some embodiments of the various procedures or aspects provided herein, the or one of the prior lines of therapy includes an IMiD. In some embodiments, the IMiD is or includes pomalidomide. In some embodiments, the IMiD is or includes lenalidomide. In some embodiments, the subject has received prior treatment comprising a combination of lenalidomide and pomalidomide.

[0011] In some embodiments of the various procedures or aspects presented here, one or more of the preceding lines of therapy include an anti-CD38 antibody. In some embodiments, the anti-CD38 antibody is daratumumab and / or isatuximab.

[0012] In some embodiments of the various procedures or aspects presented here, one or more of the preceding lines of therapy include a proteasome inhibitor. In some embodiments, the proteasome inhibitor is bortezomib, carfilzomib, ixazomib, or any combination thereof.

[0013] In some embodiments of the various procedures or aspects provided herein, the subject has further received bridge therapy. In certain embodiments, the bridge therapy is at the physician's discretion. In some embodiments, the bridge therapy includes pomalidomide, bortezomib, dexamethasone, daratumumab, or any combination thereof. In some embodiments, the bridge therapy includes pomalidomide, bortezomib, and dexamethasone. In other embodiments, the bridge therapy includes daratumumab, pomalidomide, and dexamethasone. In some embodiments, the subject has received the bridge therapy from approximately every 20 days to approximately every 30 days. In some embodiments, the subject has received the bridge therapy approximately every 21 days. In some embodiments, the subject has received the bridge therapy approximately every 28 days.In some embodiments, the subject has received at least one, two, three, four or more cycles of bridging therapies.

[0014] In some embodiments of the various procedures or aspects provided herein, the subject has further received lymphodepletion therapy. In some embodiments, the lymphodepletion therapy includes daily administration of cyclophosphamide and / or fludarabine. In some embodiments, the lymphodepletion therapy includes daily administration of cyclophosphamide and fludarabine. In some embodiments, the lymphodepletion therapy includes cyclophosphamide at a concentration of approximately 300 mg / m³. 2 and fludarabine at a concentration of approximately 30 mg / m³ 2 daily for 3 days.

[0015] In some embodiments of the various methods or aspects provided herein, the T-cell dose is 0.5-1.0 × 10 6-cells / kg body weight of the subject. In preferred embodiments, the T-cell dose is approximately 0.75 × 10 6 -cells / kg body weight of the subject. In some embodiments, the method involves administering the dose of T cells approximately 5 to 7 days after the start of lymphodepletion therapy. Preferably, the dose is administered as a single infusion.

[0016] In some embodiments of the various methods or aspects provided herein, the method is effective in maintaining an overall response in the subject after administration of the dose of T cells to the subject. In some embodiments, the method is effective in maintaining the overall response at a rate of approximately 70% to approximately 100%. In some embodiments, the method is effective in maintaining the overall response at a rate of approximately 74%. In some embodiments, the method is effective in maintaining the overall response at a rate of approximately 84.6%. In some embodiments, the method is effective in maintaining the overall response at a rate of approximately 99.4%.

[0017] In some embodiments of the various methods or aspects provided herein, the overall reaction comprises, in order from best to worst: (1) a stringently complete reaction; (2) a complete reaction; (3) a very good partial reaction; (4) a partial reaction; or (5) a minimal reaction. In some embodiments, the overall reaction is a stringently complete reaction. In some embodiments, the method is effective in obtaining the stringently complete reaction at a rate of about 40% to about 90%, about 50% to about 80%, about 58.2%, or about 68.8%. In other embodiments, the overall reaction is a complete reaction. In some embodiments, the method is effective in obtaining the complete reaction at a rate of about 10% to about 20%. In some embodiments, the method is effective in obtaining the complete reaction at a rate of about 14.9% or about 17.6%.In some embodiments, the overall reaction is a very good partial reaction or a partial reaction. In some embodiments, the method is effective in obtaining a stringently complete reaction or a complete reaction at a rate of about 70% to about 90%. In some embodiments, the method is effective in obtaining a stringently complete reaction or a complete reaction at a rate of about 73.1% or about 86.4%. In some embodiments, the method is effective in obtaining a stringently complete reaction, a complete reaction, or a very good partial reaction at a rate of about 80% to about 100%. In some embodiments, the method is effective in obtaining a stringently complete reaction, a complete reaction, or a very good partial reaction at a rate of about 81.3% or about 96.0%.In some embodiments, the method is effective in maintaining a minimal response. In some embodiments, the method is effective in maintaining a minimal residual disease negative. In some embodiments, the method is effective in maintaining a minimal residual disease negative at a rate of approximately 50% to approximately 80%. In some embodiments, the method is effective in maintaining a minimal residual disease negative at a rate of approximately 60.6%. In some embodiments, the method is effective in maintaining a minimal residual disease negative at a rate of approximately 71.6%.

[0018] In some embodiments of the various methods or aspects provided herein, the method is effective in further maintaining progression-free 12-month survival for at least approximately 60% to approximately 100% of subjects. In some embodiments, the method is effective in further maintaining progression-free 12-month survival for at least approximately 69.4% to approximately 81.1% of subjects. In some embodiments, the method is effective in further maintaining progression-free 12-month survival for at least approximately 84.1% to approximately 93.4% of subjects. In some embodiments, the method is effective in maintaining progression-free 12-month survival for at least approximately 75.9% of subjects. In some embodiments, the method is effective in maintaining progression-free 12-month survival for at least approximately 89.7% of subjects.

[0019] In some embodiments of the various methods or aspects provided herein, the time to the first total reaction or the first minimal reaction ranges from approximately 0.9 to approximately 11.1 months. In some embodiments, the median time to the first total reaction or the first minimal reaction is approximately 2.1 months.

[0020] In some embodiments of the various methods or aspects provided herein, the time to best overall response or best minimum response is approximately 1.1 to approximately 18.6 months. In some embodiments, the median time to best overall response or best minimum response is approximately 6.4 months. In some embodiments, the median time to best overall response or best minimum response is approximately 6.5 months.

[0021] In some embodiments of the various methods or aspects provided herein, the method further includes treating the subject for an adverse event following administration of the T-cell dose. In some embodiments, the method includes administering treatment to the subject to alleviate the adverse event. In some embodiments, the adverse event includes a hematological adverse event, a non-hematological adverse event, a treatment-related adverse event, or any combination thereof. In some embodiments, the non-hematological adverse event includes an infection other than an infection and / or a non-hematological adverse event other than an infection.In some embodiments, the adverse event includes neutropenia, thrombocytopenia, anemia, lymphopenia, upper respiratory tract infection, nasopharyngitis, sinusitis, rhinitis, tonsillitis, pharyngitis, laryngitis, pharyngotonsillitis, COVID-19, COVID-19 pneumonia, asymptomatic COVID-19, neutropenic sepsis, progressive multifocal leukoencephalopathy, septic shock, respiratory failure, pulmonary embolism, lower respiratory tract / lung infection, pneumonia, bronchitis, nausea, hypogammaglobulinemia, diarrhea, fatigue, headache, constipation, hypokalemia, asthenia, peripheral edema, decreased appetite, peripheral sensory neuropathy, back pain, arthralgia, pyrexia, dyspnea, insomnia, or any combination thereof. In some embodiments, the undesired event is a grade 3 / 4 undesired event.In some embodiments, the undesired event lasts longer than approximately 30 days or approximately 60 days.

[0022] In some embodiments of the various methods or aspects provided herein, the method further includes treating the subject for a second primary malignancy after administration of the T-cell dose. In some embodiments, the method includes administering treatment to the subject to alleviate the second primary malignancy. In some embodiments, the second primary malignancy includes a cutaneous / non-invasive malignancy, a hematological malignancy, a non-cutaneous / invasive malignancy, or any combination thereof.In some embodiments, the second primary malignancy includes basal cell carcinoma, Bowen's disease, squamous cell carcinoma of the lip, malignant melanoma, malignant melanoma in situ, squamous cell carcinoma of the skin, acute myeloid leukemia, a myelodysplastic syndrome, peripheral T-cell lymphoma, angiosarcoma, invasive lobular breast carcinoma, pleomorphic malignant fibrous histiocytoma, renal cell carcinoma, tonsil carcinoma, or any combination thereof.

[0023] In some embodiments of the various procedures or aspects provided herein, the adverse event or second primary malignancy occurs in the subject at a rate comparable to the rate of the same adverse event or second primary malignancy occurring in a subject undergoing standard treatment.

[0024] In some embodiments of the various methods or aspects provided herein, the method further includes treating the subject for a CAR-T-associated adverse event following administration of the T-cell dose. In some embodiments, the method includes administering treatment to the subject to alleviate the CAR-T-associated adverse event. In some embodiments, the CAR-T-associated adverse event includes cytokine release syndrome (CRS) and / or neurotoxicity.

[0025] In some embodiments of the various methods or aspects provided herein, the CAR-T-associated adverse event is a CRS. In some embodiments, the CRS occurs in the subject at a rate of approximately 60% to approximately 90%. In some embodiments, the CRS occurs in the subject at a rate of approximately 76.1%. In some embodiments, the maximum toxicity grade of the CRS is Grade 1, Grade 2, or Grade 3. In some embodiments, the maximum toxicity grade of the CRS is Grade 1. In some embodiments, the maximum toxicity grade of Grade 1 occurs in the subject at a rate of approximately 52.8%. In some embodiments, the maximum toxicity grade of the CRS is Grade 2. In some embodiments, the maximum toxicity grade of Grade 2 occurs in the subject at a rate of approximately 22.2%. In some embodiments, the maximum toxicity grade of the CRS is Grade 3.In some embodiments, the maximum toxicity grade of Grade 3 occurs in the subject at a rate of approximately 1.1%. In some embodiments, the time to first onset of CRS ranges from approximately 1 to approximately 23 days. In some embodiments, the median time to first onset of CRS is approximately 8 days. In some embodiments, the duration of CRS ranges from approximately 1 to approximately 17 days. In some embodiments, the median duration of CRS is approximately 3 days. In some embodiments of the various procedures or aspects provided herein, treatment of the adverse event includes tocilizumab, oxygen, a corticosteroid, a vasopressor, or any combination thereof.

[0026] In some embodiments of the various methods or aspects provided herein, the CAR-T-associated adverse event is neurotoxicity. In some embodiments, neurotoxicity includes an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, treatment-related adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof. In some embodiments, neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated symptom.

[0027] In some embodiments of the various methods or aspects provided herein, immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of approximately 4.5%. In some embodiments, the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1 or Grade 2. In some embodiments, the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1. In some embodiments, the maximum toxicity grade 1 occurs in the subject at a rate of approximately 3.4%. In other embodiments, the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 2. In some embodiments, the maximum toxicity grade 2 occurs in the subject at a rate of approximately 1.1%.In some embodiments, the time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from approximately 6 to approximately 15 days. In some embodiments, the median time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom is approximately 9.5 days. In some embodiments, the duration of immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from approximately 1 to approximately 6 days. In some embodiments, the duration of immune effector cell-associated neurotoxicity syndrome or associated symptom is approximately 2 days. In some embodiments, treatment of the adverse event includes a corticosteroid and / or tocilizumab.

[0028] In some embodiments of the various procedures or aspects provided herein, the neurotoxicity is CAR-T cell neurotoxicity. In some embodiments, the CAR-T cell neurotoxicity occurs in the subject at a rate of approximately 17.0%. In some embodiments, the CAR-T cell neurotoxicity includes grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, an adverse event induced by movement and neurocognitive treatment, or any combination thereof. In some embodiments, the CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity occurring in the subject at a rate of approximately 2.3%. In some embodiments, the CAR-T cell neurotoxicity is grade 5 neurotoxicity. In some embodiments, the CAR-T cell neurotoxicity is cranial nerve palsy.In some embodiments, cranial nerve palsy occurs in the subject at a rate of approximately 9.1%. In some embodiments, the cranial nerve palsy is grade 2 or grade 3. In some embodiments, the cranial nerve palsy is grade 2, occurring in the subject at a rate of approximately 8.0%. In some embodiments, the cranial nerve palsy is grade 3, occurring in the subject at a rate of approximately 1.1%. In some embodiments, the time to onset of cranial nerve palsy after administration of the T-cell dose to the subject ranges from approximately 17 days to approximately 60 days. In some embodiments, the median time to onset of cranial nerve palsy after administration of the T-cell dose to the subject is approximately 21 days.In some embodiments, cranial nerve palsy affects cranial nerves III, V, or VII. In some embodiments, the duration of cranial nerve palsy ranges from about 15 days to about 262 days. In some embodiments, the median duration of cranial nerve palsy is about 77 days. In some embodiments, the treatment includes a corticosteroid. In some embodiments, CAR T-cell neurotoxicity is peripheral neuropathy. In some embodiments, peripheral neuropathy occurs in the subject at a rate of about 2.8%. In some embodiments, the peripheral neuropathy is grade 1. In some embodiments, grade 1 peripheral neuropathy occurs at a rate of about 1.1%. In some embodiments, the peripheral neuropathy is grade 2. In some embodiments, grade 2 peripheral neuropathy occurs at a rate of about 1.1%. In some embodiments, the peripheral neuropathy is grade 3.In some embodiments, grade 3 peripheral neuropathy occurs at a rate of approximately 0.6%. In some embodiments, CAR-T cell neurotoxicity is an adverse event induced by movement and neurocognitive treatment. In some embodiments, the adverse event induced by movement and neurocognitive treatment is grade 1. In some embodiments, the grade 1 movement and neurocognitive treatment-related adverse event occurs in the subject at a rate of approximately 0.6%.

[0029] In some embodiments of the various methods or aspects provided herein, CD3+ cells encompassing the CAR in the subject's blood reach their peak approximately 13 days after administration of the T cells to the subject. In some embodiments, the CD3+ cells encompassing the CAR in the subject's blood reach a peak concentration of approximately 1523 cells / µl. In some embodiments, CD3+ cells encompassing the CAR in the subject's blood remain detectable from approximately 13 days to approximately 631 days after administration of the T cells to the subject. In some embodiments, the CD3+ cells encompassing the CAR in the subject's blood remain detectable for a median of approximately 57 days after administration of the T cells to the subject. In some embodiments, the AUC is 0-28 of CD3+ cells encompassing the CAR in the subject's blood, at a mean of approximately 12,504 cells / µL.

[0030] In some embodiments of the various methods or aspects provided herein, the first VHH domain comprises a CDR1, a CDR2 and a CDR3 of the VHH domain comprising the amino acid sequence of SEQ ID NO: 2, and the second VHH domain comprises a CDR1, a CDR2 and a CDR3 of the VHH domain comprising the amino acid sequence of SEQ ID NO: 4. In some embodiments, the first VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 18, a CDR2 comprising the amino acid sequence of SEQ ID NO: 19, a CDR3 comprising the amino acid sequence of SEQ ID NO: 20, and the second VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a CDR2 comprising the amino acid sequence of SEQ ID NO: 22, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 23. In some embodiments, the first VHH domain comprises the amino acid sequence of SEQ ID NO: 2 and the second VHH domain comprises the amino acid sequence of SEQ ID NO: 4.In some embodiments, the first VHH domain is located at the N-terminus of the second VHH domain. In other embodiments, the first VHH domain is located at the C-terminus of the second VHH domain. In some embodiments, the first VHH domain is located via a linker comprising the amino acid sequence of SEQ ID NO: 3. In some embodiments, the transmembrane domain is derived from a molecule selected from the group consisting of CD8α, CD4, CD28, CD137, CD80, CD86, CD152, and PD1. In some embodiments, the transmembrane domain is derived from CD8α and comprises the amino acid sequence of SEQ ID NO: 6. In some embodiments, the intracellular signaling domain comprises a primary intracellular signaling domain of an immune effector cell. In some embodiments, the primary intracellular signaling domain is derived from CD3ζ and comprises the amino acid sequence of SEQ ID NO: 8.In some embodiments, the intracellular signaling domain comprises a costimulatory signaling domain. In some embodiments, the costimulatory signaling domain is derived from a costimulatory molecule selected from the group consisting of CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, ligands of CD83, and any combination thereof. In some embodiments, the costimulatory signaling domain comprises a cytoplasmic domain of CD137 comprising the amino acid sequence of SEQ ID NO: 7. In some embodiments, the CAR further comprises a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the joint domain is derived from CD8α, comprising the amino acid sequence of SEQ ID NO: 5.In some embodiments, the CAR further comprises a signal peptide located at the N-terminus of the polypeptide. In some embodiments, the signal peptide is derived from CD8α, comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the CAR comprises the amino acid sequence of SEQ ID NO: 17.

[0031] In some aspects, methods for treating a subject with multiple myeloma are provided herein, wherein the method involves administering to the subject a dose of T cells comprising a chimeric antigen receptor (CAR) comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of the B-cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD. In some embodiments, the administration of the dose of T cells reduces the risk of disease progression or death in the subject.In some embodiments, the risk of disease progression or death is reduced compared to treatment with daratumumab pomalidomide dexamethasone (DPd) or pomalidomide bortezomib dexamethasone (PVd). In some embodiments, the risk of disease progression or death is reduced compared to standard of care (SOC) therapy, which includes administration of daratumumab pomalidomide dexamethasone (DPd) or pomalidomide bortezomib dexamethasone (PVd). In some embodiments, the risk of disease progression or death is reduced compared to administration of the Ide-Cel treatment. In some embodiments, the subject has an approximately 60% to approximately 75% reduced risk of disease progression or death. In some embodiments, the subject has an approximately 74% reduced risk of disease progression or death.

[0032] In some aspects, methods for treating a subject with multiple myeloma are provided herein, wherein the method comprises administering to the subject a dose of T cells comprising a chimeric antigen receptor (CAR) comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of the B-cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of therapy, including immunomodulatory drug (IMiD) therapy, and is refractory to the IMiD, and wherein administering the dose of T cells is more effective than administering DPd or PVd treatment in obtaining a very good partial response (VGPR) or better in the subject. In some embodiments, the VGPR or better after administration of the treatment is approximately 81.3%.In some embodiments, the VGPR or better after administration of DPd or PVd is approximately 45.5%. In some embodiments, the VGPR or better after administration of standard treatment (SOC) therapy comprising the administration of either DPd or PVd is approximately 45.5%. In some embodiments, the treatment is more effective in obtaining a stringent complete response (sCR) in the subject compared with the administration of DPd or PVd alone. In some embodiments, the sCR after administration of the treatment is approximately 58.2%. In some embodiments, the sCR after administration of DPd or PVd is approximately 15.2%.

[0033] Another aspect of the disclosure is a method for treating a subject with multiple myeloma, comprising administering a dose of T cells comprising a chimeric antigen receptor (CAR) to the subject, comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of the B-cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD.

[0034] In some embodiments, the subject has a reduced risk of developing a CAR-T-associated adverse event, optionally including a cytokine release syndrome (CRS) and / or neurotoxicity.

[0035] In some embodiments, the CAR-T-associated adverse event is a CRS, wherein the CRS may further optionally occur in the subject at a rate of 60% to about 90% or at a rate of about 76.1%, and wherein the maximum toxicity grade of the CRS may further optionally be Grade 1, Grade 2 or Grade 3.

[0036] In some embodiments, the CAR-T-associated adverse event is a CRS, wherein the CRS may further optionally occur in the subject at a rate of about 60% to about 90% or at a rate of about 76.1%, wherein further optionally the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, wherein further optionally the maximum toxicity grade of the CRS is Grade 1, wherein optionally the maximum toxicity grade of Grade 1 occurs in the subject at a rate of about 52.8%.

[0037] In some embodiments, the CAR-T-associated adverse event is a CRS, wherein the CRS may further optionally occur in the subject at a rate of about 60% to about 90% or at a rate of about 76.1%, wherein further optionally the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, wherein further optionally the maximum toxicity grade of the CRS is Grade 2, wherein optionally the maximum toxicity grade of Grade 2 occurs in the subject at a rate of about 22.2%.

[0038] In some embodiments, the CAR-T-associated adverse event is a CRS, wherein the CRS may further optionally occur in the subject at a rate of about 60% to about 90% or at a rate of about 76.1%, wherein further optionally the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, wherein further optionally the maximum toxicity grade of the CRS is Grade 3, wherein optionally the maximum toxicity grade of Grade 3 occurs in the subject at a rate of about 1.1%;

[0039] In some embodiments, the CAR-T-associated adverse event is a CRS, wherein the CRS may further optionally occur in the subject at a rate of about 60% to about 90% or at a rate of about 76.1%, wherein further optionally the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, wherein further optionally the time to first onset of the CRS is in the range of about 1 to about 23 days, wherein optionally the time to first onset of the CRS is in a median of about 8 days.

[0040] In one embodiment, the CAR-T-associated adverse event is a CRS, wherein the CRS may further optionally occur in the subject at a rate of about 60% to about 90% or at a rate of about 76.1%, wherein further optionally the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, wherein further optionally the duration of the CRS is in the range of about 1 to about 17 days, wherein optionally the duration of the CRS is in a median of about 3 days.

[0041] In some embodiments, the CAR-T-associated adverse event is a CRS, wherein the CRS further optionally occurs in the subject at a rate of about 60% to about 90% or at a rate of about 76.1%, wherein further optionally the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3, wherein the method further optionally comprises administering tocilizumab, oxygen, a corticosteroid, or a vasopressor combination thereof to the subject.

[0042] In some embodiments, the CAR-T-associated adverse event is neurotoxicity, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, treatment-related adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated neurotoxicity syndrome.

[0043] In some embodiments, the CAR-T-associated adverse event is neurotoxicity, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, treatment-related adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated symptom, further optionally comprising the immune effector cell-associated neurotoxicity syndrome or associated symptom occurring in the subject at a rate of approximately 4.5%.

[0044] In some embodiments, the CAR-T-associated adverse event is neurotoxicity, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, treatment-related adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated symptom, optionally comprising the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom being Grade 1 or Grade 2, optionally comprising the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom being Grade 1.where, where furthermore, the maximum toxicity level may occur at a rate of approximately 3.4%, grade 1, or where, where applicable, the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is grade 2, where furthermore, where applicable, the maximum toxicity level of grade 2 occurs in the subject at a rate of approximately 1.1%.

[0045] In some embodiments, the CAR-T-associated adverse event is neurotoxicity, wherein the neurotoxicity optionally comprises an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, treatment-related adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, wherein optionally the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated symptom, wherein further optionally the time to onset of the immune effector cell-associated neurotoxicity syndrome or associated symptom is in the range of about 6 to about 15 days, wherein optionally the time to onset of the immune effector cell-associated neurotoxicity syndrome or associated symptom is in the median of about 9.5 days;

[0046] In some embodiments, the CAR-T-associated adverse event is neurotoxicity, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, treatment-related adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated symptom, optionally comprising the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom being in the range of about 1 to about 6 days, optionally comprising the median duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom being about 2 days.

[0047] In some embodiments, the CAR-T-associated adverse event is neurotoxicity, optionally comprising an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, standard-of-care-arm adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, optionally comprising an immune effector cell-associated or associated neurotoxicity syndrome, and comprising zumab and / or an associated neurotoxicity syndrome.

[0048] Another aspect of the disclosure is a method for treating a subject with multiple myeloma, comprising administering a dose of T cells comprising a chimeric antigen receptor (CAR) to the subject, comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of the B-cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD.

[0049] In some embodiments, the subject has a reduced risk of developing a CAR-T-associated adverse event, optionally comprising CAR-T-associated adverse event, wherein optionally CAR-T-associated adverse event comprises CAR-T cell neurotoxicity, wherein optionally CAR-T cell neurotoxicity occurs in the subject at a rate of approximately 17.0%, and optionally comprising grade 3 / 4 neurotoxicity or grade 5 neurotoxicity.

[0050] In some embodiments, CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, occurring in the subject at a rate of approximately 2.3%.

[0051] In some embodiments, CAR-T cell neurotoxicity is grade 5 neurotoxicity.

[0052] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, with cranial nerve palsy occurring in the subject at a rate of approximately 9.1%.

[0053] In some embodiments, CAR T-cell neurotoxicity is cranial nerve palsy, optionally occurring in the subject at a rate of approximately 9.1%. The cranial nerve palsy is grade 2 or grade 3, optionally being grade 2 occurring in the subject at a rate of approximately 8.0%, and optionally being grade 3 occurring in the subject at a rate of approximately 1.1%.

[0054] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, wherein cranial nerve palsy may occur in the subject at a rate of approximately 9.1% until the onset of cranial nerve palsy after administration of the dose of T cells to the subject is in the range of approximately 17 days to approximately 60 days, and wherein, furthermore, the time until the onset of cranial nerve palsy after administration of the dose of T cells to the subject may have a median of approximately 21 days.

[0055] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, with the cranial nerve palsy possibly occurring in the subject at a rate of about 9.1% and affecting cranial nerve III, V or VII.

[0056] In some embodiments, CAR-T cell neurotoxicity is cranial nerve palsy, wherein cranial nerve palsy may occur in the subject at a rate of about 9.1% and the duration of cranial nerve palsy may range from about 15 days to about 262 days, and wherein the median duration of cranial nerve palsy may further be about 77 days.

[0057] In some embodiments, the CAR-T cell neurotoxicity is cranial nerve palsy, wherein, optionally, the cranial nerve palsy occurs in the subject at a rate of about 9.1%, and the procedure further comprises administering a treatment comprising a corticosteroid to the subject.

[0058] In some embodiments, CAR T-cell neurotoxicity is peripheral neuropathy, with peripheral neuropathy occurring in the subject at a rate of approximately 2.8%. In some embodiments, the peripheral neuropathy is grade 1. In some embodiments, grade 1 peripheral neuropathy occurs at a rate of approximately 1.1%. In some embodiments, the peripheral neuropathy is grade 2. In some embodiments, grade 2 peripheral neuropathy occurs at a rate of approximately 1.1%. In some embodiments, the peripheral neuropathy is grade 3. In some embodiments, grade 3 peripheral neuropathy occurs at a rate of approximately 0.6%.

[0059] In some embodiments, CAR-T cell neurotoxicity is an adverse event resulting from movement and neurocognitive treatment, wherein optionally the adverse event resulting from movement and neurocognitive treatment is grade 1, and furthermore, optionally the grade 1 movement and the adverse event resulting from neurocognitive treatment occur in the subject at a rate of about 0.6%.

[0060] Features described in the context of separate aspects and embodiments of the disclosure may be used together and / or be interchangeable. Likewise, features described in connection with a single embodiment may also be provided separately or in any suitable subcombination. All methods described herein, but expressed, may be described as corresponding uses, in particular medical uses. BRIEF DESCRIPTION OF THE DRAWINGS Fig. Figure 1 shows the expression of BCMA antigen on the surface of GC, memory, and plasmablast cells in lymph nodes, long-lived plasma cells in bone marrow (LN and MALT), and on multiple myeloma cells. BAFF-R antigen is not expressed on plasmablast cells, long-lived plasma cells, or multiple myeloma cells. TACI is expressed on memory and plasmablast cells, long-lived plasma cells, and multiple myeloma cells. CD138 is expressed only on long-lived plasma cells and multiple myeloma cells. Fig. Figure 2 shows the structure of the ciltacabtagene autoleucel CAR. Ciltacabtagene autoleucel comprises two VHH domains, in contrast to the single VL and VH domains found on various other CARs. Ciltacabtagene autoleucel includes intracellular CD137 and human CD3 zeta domains. Fig.Figure 3 shows a scheme for the production of a ciltacabtagene autoleucel CAR-encoding virus, for the transduction of the virus into a patient T cell, and subsequently for the production of CAR-T cells expressing ciltacabtagene autoleucel. Fig. Figure 4 shows a schematic study design for ciltacabtagene autoleucel CAR T cells. The patient population comprises patients with relapsed or refractory multiple myeloma who have received one to three prior lines of therapy, including an immunomodulatory drug or who are double-refractory to PI / IMiD and have prior exposure to PI, IMiD, and anti-CD38. A primary objective is to compare the efficacy and safety of ciltacabtagene autoleucel CAR T cells with physician choice between two highly effective standard-of-care therapies in the patient population described above in a randomized controlled trial (phase 3). Fig.Figure 5 is a diagram showing the disposition of the study participant in each treatment arm. Fig. Figures 6A-6C show Kaplan-Meier Intent-to-Treat analysis. Fig. Figure 6A shows progression-free survival after treatment arm. Fig. 6B shows progression-free survival after treatment arm and stratified by number of previous lines of therapy. Fig. 6C shows the overall survival after treatment. Fig. Figure 7 shows the forest plot of the subgroup analysis of progression-free survival. Abbreviations: Cilta-cel, ciltacabtagene autoleucel; DPd, daratumumab pomalidomide dexamethasone; ECOG, Eastern Cooperative Oncology Group; IMID, immunomodulatory drug; ISS, International Staging System; MM, multiple myeloma; NCI, National Cancer Institute; PI, proteasome inhibitor; PVd, pomalidomide bortezomib dexamethasone; SOC, standard of care. aHazard ratio and 95% CI from a Cox proportional hazards model with treatment as the sole explanatory variable, including only PFS events occurring > 8 weeks after randomization. A hazard ratio <1 indicates a benefit for the Cilta-cel arm. b Based on the randomization strata of the interactive web response system. c Based on serum β-2 microglobulin and albumin. d Based on subjects with measurable disease in serum. e Positive for del(17p), t(14;16), t(4;14) and / or Gain / amp(1q) by FISH testing. Protocol-defined high-risk cytogenetics refers to one of four abnormal markers. f Based on the modification of the formula for nutrition in kidney disease (MDRD). Fig. Figures 8A-8D show the comparison of PFS and OS responses in patients who received Cilta-cel in CARTITUDE-1 and in CARTITUDE-4. Fig.8A shows PFS in patients who received Cilta-cel as the study treatment in CARTITUDE-1. Fig. 8B shows PFS in patients who received Cilta-cel as the study treatment in CARTITUDE-4. Fig. 8C shows OS in patients who received Cilta-cel as the study treatment in CARTITUDE-1. Fig. 8D shows OS in patients who received Cilta-cel as the study treatment in CARTITUDE-4. DETAILED DESCRIPTION

[0061] The disclosure also provides related nucleic acids, recombinant expression vectors, host cells, cell populations, antibodies, or antigen-binding components thereof, and pharmaceutical compositions relating to the immune cells and CAR-expressing T cells of the disclosure. Dosage regimens and dosage forms, as well as procedures for treatment with the CAR-T cells, are also provided.

[0062] Several aspects and embodiments of the disclosure are described below with reference to examples for illustrative purposes only. It is understood that numerous specific details, relationships, and procedures are set forth to provide a complete understanding of the disclosure. However, a person skilled in the art will readily recognize that the disclosure can be practiced without one or more of the specific details or with different procedures, protocols, reagents, cell lines, and animals. The present disclosure is not limited by the illustrated sequence of actions or events, since some actions may occur in different sequences and / or concurrently with other actions or events. Furthermore, not all illustrated actions, steps, or events are required to implement a methodology according to the present disclosure.

[0063] Unless otherwise defined, all terms, designations, and other scientific terms or terminology used herein shall have the meanings generally understood by a person skilled in the art to which this disclosure refers. In some cases, terms with generally understood meanings are defined here for clarity and / or for ready reference, and the inclusion of such definitions here should not necessarily be interpreted as representing a material difference from what is generally understood in the prior art. It is further understood that terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant field and / or as defined differently here.

[0064] The term "approximately" or "about" implies that the value lies within a statistically significant range. Such a range may be of a certain order of magnitude, preferably within 50%, particularly preferably within 20%, particularly preferably within 10%, and even more preferably within 5% of a given value or range. The permissible variation encompassed by the term "approximately" or "about" depends on the specific system under investigation and can be readily understood by a person skilled in the art.

[0065] The term "antibody" includes monoclonal antibodies (including full-length, four-chain antibodies or full-length, heavy-chain-only antibodies possessing an immunoglobulin FC region), antibody compositions with polyepitopic specificity, multispecific antibodies (e.g., bispecific antibodies, diabodies, and single-chain molecules), and antibody fragments (e.g., Fab, F(ab')2, and FV). The term "immunoglobulin" (Ig) is used interchangeably with "antibody" here. Antibodies considered here include single-domain antibodies, such as heavy-chain-only antibodies.

[0066] The term "heavy-chain-only antibody" or "HCAb" refers to a functional antibody that includes heavy chains but lacks the light chains typically found in four-chain antibodies. Camelids (such as camels, llamas, or alpacas) are known to produce HCAbs.

[0067] The term "single-domain antibody" or "sdAb" refers to a single antigen-binding polypeptide with three complementary target regions (CDRs). The sdAb alone is able to bind to the antigen without pairing with a corresponding CDR-containing polypeptide. In some cases, single-domain antibodies are genetically engineered from camelid heavy chain antibodies (HCAbs), and their variable heavy chain domains are referred to as "VHHs." Some VHHs may also be known as "nanobodies." A KamelidsdAb is one of the smallest known antigen-binding antibody fragments (see e.g. Hamers-Casterman et al., Nature 363:446-8 (1993); Greenberg et al., Nature 374:168-73 (1995); Hassanzadeh-Ghassabeh et al., nanomedicine (Lond), 8:1013-26 (2013)).A basic VHH has the following structure from the N-terminus to the C-terminus: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, where FR1 to FR4 each refer to scaffold regions 1 to 4 and where CDR1 to CDR3 refer to the complementarity determination regions 1 to 3.

[0068] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domains of the antibody's heavy or light chain. The variable domains of the heavy chain and light chain can be designated "VH" and "VL," respectively. These domains are generally the most variable parts of the antibody (compared to other antibodies of the same class) and contain the antigen-binding sites. Only heavy-chain antibodies from camelid species possess a single variable region of the heavy chain, designated as the "VHH" domain. VHH is thus a special type of variable region.

[0069] The term "variable" refers to the fact that certain segments of the variable domains differ significantly in sequence among antibodies. The V domain (i.e., variable domain) mediates antigen binding and defines the specificity of a given antibody for its specific antigen. However, variability is not uniformly distributed across the entire range of variable domains. Instead, it is concentrated in three segments called hypervariable regions (HVRs), found in both the light chain and heavy chain variable domains. The more highly conserved portions of variable domains are called scaffold regions (FRs).The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a β-sheet configuration, connected by three HVRs that form loops linking and partially forming part of the β-sheet structure. The HVRs in each chain are held together by the closely spaced FR regions and contribute to the formation of the antibody's antigen-binding site (along with the HVRs from the other chain if the antibody is not a sdAb or HCAb) (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institutes of Health, Bethesda, MD (1991)). The constant domains are not directly involved in antibody binding to an antigen but exhibit various effector functions, such as the antibody's involvement in antibody-dependent cellular toxicity.

[0070] The terms “antibody fragment,” “antibody fragment,” “functional antibody fragment,” and “antigen-binding portion” are used interchangeably here to mean one or more fragments or parts of an antibody that retain the ability to bind specifically to an antigen (see generally Holliger et al., Nat. Biotech., 23(9): 1126–1129 (2005)). The antigen recognition unit of the CARs, encoded by the nucleic acid sequences disclosed herein, may contain any BCMA-binding antibody fragment. The antibody fragment may conveniently include, for example, one or more CDRs, the variable region (or parts thereof), the constant region (or parts thereof), or combinations thereof. Examples of antibody fragments include, but are not limited to,(i) a Fab fragment, which is a monovalent fragment consisting of the VL, VH, CL and CHI domains; (II) an F(ab')2 fragment, which is a divalent fragment comprising two Fab fragments linked by a disulfide bridge at the joint region; (III) an FV fragment consisting of the VL and VH domains of a single arm of an antibody; (iv) a single-chain FV (scFv) which is a monovalent molecule consisting of the two domains of the FV fragment (i.e., VL and VH) linked by a synthetic linker that allows the two domains to be synthesized as a single polypeptide chain (see, e.g., Bird et al., Science, 242: 423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA, 85: 5879-5883 (1988); and Osbourn et al., Nat.Biotechnol, 16: 778 (1998)) and (v) a diabody, which is a dimer of polypeptide chains, wherein each polypeptide chain comprises a VH linked to a VL by a peptide linker too short to allow pairing between the VH and the VL on the same polypeptide chain, thereby driving pairing between the complementary domains on different VH-VL polypeptide chains to generate a dimeric molecule with two functional antigen-binding sites. Antibody fragments are known in the prior art and are described in more detail, for example, in US patent application 2009 / 0093024 A1.

[0071] Within the scope of the present invention, the terms “specifically binds”, “specifically recognizes” or “specific for” refer to measurable and reproducible interactions, such as the binding between a target and an antigen-binding protein (such as a CAR or a VHH) that is determinative for the presence of the target in the presence of a heterogeneous population of molecules, including biological molecules.

[0072] The term "specificity" refers to the selective recognition of an antigen-binding protein (such as a CAR or a VHH) for a specific epitope of an antigen. Natural antibodies, for example, are monospecific.

[0073] A chimeric antigen receptor, or CAR, is an artificially engineered hybrid protein or polypeptide containing the antigen-binding domains of an antibody (or antibody fragment) linked to T-cell signaling domains. Properties of CARs may include their ability to redirect T-cell specificity and reactivity to a selected target in a non-MHC-restricted manner, thereby exploiting the antigen-binding properties of monoclonal antibodies. This non-MHC-restricted antigen recognition gives CAR-expressing T cells the ability to recognize antigens independently of antigen processing, bypassing a key mechanism of tumor excretion. Furthermore, when expressed in T cells, CARs advantageously do not dimerize with the α- and β-chains of the endogenous T-cell receptor (TCR).T cells expressing a CAR are referred to here as CAR-T cells, CAR-T cells, or CAR-modified T cells, and these terms are used interchangeably. The cell can be genetically modified to stably express an antibody-binding domain on its surface, conferring novel antigen specificity that is MHC-independent. “BCMA-CAR” refers to a CAR with an extracellular binding domain specific for BCMA. “BI-Epitope-CAR” refers to a CAR with an extracellular binding domain specific for two different epitopes on BCMA.

[0074] Ciltacabtagene autoleucel (“Cilta-cel”) is a chimeric antigen receptor T-cell (CAR-T) therapy comprising two B-cell maturation antigen (BCMA)-targeting VHH domains designed to confer BCMA avidity. Cilta-cel can include T lymphocytes transduced with the ciltacabtagene autoleucel CAR, a CAR encoded by a lentiviral vector. The CAR targets the human B-cell maturation antigen (BCMA CAR). A diagram of the lentiviral vector encoding Cilta-cel CAR is shown in Fig. 2. The amino acid sequence of Cilta-cel-CAR is the amino acid sequence of SEQ ID NO: 17.

[0075] The terms "express" and "expression" mean that the information in a gene or DNA sequence can be produced or lead to its production. For example, expression can occur in the form of protein production through the activation of cellular functions involved in the transcription and translation of a corresponding gene or DNA sequence. A DNA sequence is expressed in or by a cell to form an "expression product," such as a protein. The expression product itself, e.g., the resulting protein, can also be "expressed" by the cell. An expression product can be characterized as intracellular, extracellular, or transmembrane.

[0076] The terms "treat" or "treatment" refer to any therapeutic intervention aimed at slowing or reducing an undesirable physiological change or disease, or at providing a beneficial or desired clinical outcome during treatment. Beneficial or desired clinical outcomes include relief of symptoms, reduction of disease severity, stabilization (i.e., non-worsening) of the disease state, delay or slowing of disease progression, alleviation or palliation of the disease state, and / or remission (wholly or partially), whether demonstrable or undetectable. "Treatment" can also mean that survival is prolonged compared to the expected survival if a subject does not receive treatment.Subjects requiring treatment include those already struggling with the undesired physiological change or disease, as well as those susceptible to it. Treatment may involve a treatment agent, also referred to here as a "medication" or "drug," which can be used to achieve the positive or desired clinical outcome of interest through its action. Treatment agents or drugs can be administered to a subject in many ways, including at least intravenously and orally. The term "intravenous," in the context of administering treatment agents or drugs, refers to administering these treatment agents or drugs within one or more veins.The term "oral" in connection with the administration of treatments or medications refers to the administration of these treatments or medications via an oral passage such as the mouth.

[0077] Within the scope of the present invention, the term "subject" refers to an animal. The terms "subject" and "patient" may be used interchangeably here with respect to a subject. Therefore, a "subject" includes a human being treated for a disease or for the prevention of a disease. The methods described herein can be used to treat an animal subject belonging to any classification. Examples of such animals are mammals. Mammals include, among others, mammals of the order Rodentia, such as mice and hamsters, and mammals of the order Logomorpha, such as rabbits. The mammals may belong to the order Carnivora, including Felidae (cats) and Canidae (dogs). The mammals may belong to the order Artiodactyla, including Bovidae (cows) and Suidae (pigs), or to the order Perssodactyla, including Equidae (horses).The mammals can be primates of the order cephaloids or simoids (monkeys) or anthropoids of the order anthropoids (humans and monkeys). In some versions, the mammal is a human.

[0078] The term "effective," when applied to dose or quantity, refers to the amount of a compound or pharmaceutical composition sufficient to produce a desired activity when administered to a subject who requires it. It should be noted that when administering a combination of active ingredients, the effective amount of the combination may or may not include amounts of each component that would have been effective if administered individually. The precise amount required varies from subject to subject, depending on the subject's species, age, and general condition, the severity of the condition being treated, the specific medication(s) used, the route of administration, and other factors.

[0079] The term “pharmaceutically acceptable,” as used in connection with compositions described herein, refers to molecular units and other constituents of such compositions that are physiologically acceptable and typically do not cause adverse reactions when administered to a mammal (e.g., a human). Preferably, the term “pharmaceutically acceptable” means that it is approved by a federal or state regulatory agency or is listed in the U.S. Pharmacopoeia or other generally accepted pharmacopoeias for use in mammals, and especially in humans.

[0080] The term "line of therapy," as used in the context of the treatment procedures described herein, refers to one or more cycles of a planned treatment program, which may consist of one or more planned cycles of single-agent or combination therapy, as well as a sequence of treatments administered in a planned manner. For example, a planned treatment approach of induction therapy followed by autologous stem cell transplantation followed by maintenance therapy constitutes a line of therapy. A new line of therapy is considered to have begun when a planned course of therapy has been changed to other treatment agents or drugs (alone or in combination) due to disease progression, relapse, or toxicity.A new line of therapy is also accepted if a planned observation period outside of therapy was interrupted by additional treatment of the disease.

[0081] The term "refractory," as used herein in connection with treatment with a particular treatment agent or drug, refers to diseases or patients who do not respond to the treatment agent or drug. The term "refractory myeloma" refers to a disease that does not respond during primary or salvage therapy or progresses within 60 days of the last therapy.

[0082] The term “non-response disease” refers either to the failure to achieve a minimal response or to the development of a progressive disease during therapy.

[0083] The term "hazard ratio" refers to a measure of the relative rate of progression to an endpoint compared to a control group. In outcome-based clinical trials, a reduction in the hazard ratio for a treatment arm compared to the control indicates that the treatment used in the treatment arm reduces the risk of the endpoint, in the case of the studies described here, disease progression or death. Preferably, the hazard ratio is calculated per stratified constant piecewise weighted log-rank test.

[0084] The terminology used here serves only to describe certain aspects or forms and is not intended to be restrictive. As used herein, the indefinite articles "ein", "eine", and "das" should be understood to include the plural unless the context clearly indicates otherwise.

[0085] In the disclosure, various aspects and embodiments of the disclosure may consistently be presented in a range format. It is understood that the description in range format serves only for simplicity and brevity and should not be interpreted as an inflexible limitation of the scope of protection afforded by the disclosure. Therefore, the description of a range should expressly disclose all possible subranges as well as individual numerical values ​​within that range. For example, the description of a range such as 1 to 6 should be considered to disclose subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numbers within that range, for example, 1, 2, 2, 7, 3, 4, 5, 5, 3, and 6.As another example, a range like 95-99% identity encompasses something with 95%, 96%, 97%, 98%, or 99% identity and includes subranges like 96-99%, 96-98%, 96-97%, 97-99%, 97-98%, and 98-99% identity. This holds true regardless of the range's width. vectors

[0086] Polynucleotide sequences encoding the CARs described in the present application can be obtained using standard recombinant techniques. Desired polynucleotide sequences can be isolated and sequenced from antibody-producing cells such as hybridoma cells. Alternatively, polynucleotides can be synthesized using nucleotide synthesizers or PCR techniques.

[0087] The disclosure also provides a vector comprising the nucleic acid sequence encoding the CARs disclosed herein. The vector may be, for example, a plasmid, a cosmid, a viral vector (e.g., retroviral or adenoviral), or a phage. Suitable vectors and methods for vector preparation are well known in the prior art (see, e.g., Sambrook et al. and Ausubel et al.).

[0088] In addition to the nucleic acid sequences encoding the CARs disclosed herein, the vector preferably includes expression control sequences, such as promoters, enhancers, polyadenylation signals, transcription terminators, internal ribosome entry sites (IRES), and the like, which ensure the expression of the nucleic acid sequence in a host cell. Exemplary expression control sequences are known in the prior art and are described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology, Vol. 185, Academic Press, San Diego, California (1990).

[0089] In some embodiments, the vector includes a promoter. A variety of promoters, recognized by a variety of potential host cells, are well known. The selected promoter can be operatively linked to cistron DNA encoding the CARs disclosed herein by removing the promoter from the starting DNA via restriction enzyme digestion and inserting the isolated promoter sequence into the vector of the present application. A variety of promoters, including constitutive, inducible, and repressible promoters, from a variety of different sources are well known in the prior art. Representative sources of promoters include, for example, viruses, mammals, insects, plants, yeast, and bacteria. Suitable promoters from these sources are readily available or can be synthetically produced, based on sequences, and are publicly available, for example, from repositories such as the ATCC, as well as other commercial or individual sources.Promoters can be unidirectional (i.e., initiating transcription in one direction) or bidirectional (i.e., initiating transcription in a 3' or 5' direction). Non-restrictive examples of promoters include the bacterial expression system T7, the bacterial expression system pBAD (araA), the cytomegalovirus (CMV) promoter, the SV40 promoter, and the RSV promoter. Examples of inducible promoters include the Tet system (US patents 5,464,758 and 5,814,618), the ecdysone-inducible system (No et al., Proc. Natl. Acad. Sci., 93: 3346-3351 (1996)), the T-REX-™ system (Invitrogen, Carlsbad, CA), the LACSWITCH™ system (Stratagene, San Diego, CA), and the CRE-ERT-tamoxifen-inducible recombinase system (Indra et al., NUC. Acid. Res., 27: 4324-4327 (1999); Nuc. Acid. Res., 28: e99 (2000); US patent 7,112,715; and Kramer & Fussenegger, Methods Mol. Biol, 308: 123-144 (2005)).

[0090] In some embodiments, the vector includes an “enhancer.” The term “enhancer,” as used herein, refers to a DNA sequence that enhances the transcription of, for example, a nucleic acid sequence to which it is operably linked. Enhancers may be located many kilobases away from the coding region of the nucleic acid sequence and may mediate the binding of regulatory factors, patterns of DNA methylation, or changes in DNA structure. A large number of enhancers from a variety of different sources are well known in the prior art and are available as or within cloned polynucleotides (e.g., from repositories such as the ATCC as well as other commercial or individual sources). A number of polynucleotides that include promoters (such as the commonly used CMV promoter) also include enhancer sequences. Enhancers may be located upstream, within, or downstream of coding sequences.The term “Ig enhancer” refers to enhancer elements derived from enhancer regions mapped within the immunoglobulin (Ig) locus. Such Ig enhancers include, for example, the 5' heavy chain (mu) enhancers, 5' light chain (kappa) enhancers, kappa and mu intronic enhancers, and 3' enhancers (see generally Paul WE (ed), Fundamental Immunology, 3rd edition, Raven Press, New York (1993), pages 353–363; and US Patent 5,885,827).

[0091] In some embodiments, the vector comprises a “selectable marker gene.” The term “selectable marker gene,” as used herein, refers to a nucleic acid sequence that makes it possible to select cells expressing the nucleic acid sequence specifically for or against it in the presence of a corresponding selective agent. Suitable selectable marker genes are known in the prior art and are described, for example, in international patent applications WO 1992 / 08796 and WO 1994 / 28143; Wigler et al., Proc. Natl. Acad. Sci. USA, 77: 3567 (1980); O'Hare et al., Proc. Natl. Acad. Sci. USA, 78: 1527 (1981); Mulligan & Berg, Proc. Natl. Acad. Sci. USA, 78: 2072 (1981). Colberre-Garapin et al., J. Mol. Biol., 150: 1 (1981); Santerre et al., Gene, 30:147 (1984); Kent et al., Science, 237: 901-903 (1987); Wigler et al., Cell, IP. 223 (1977); Szybalska & Szybalski, Proc. Natl. Acad. Sci. USA, 48: 2026 (1962); Lowy et al., Cell, 22: 817 (1980); and US patents 5,122,464 and 5,770,359.

[0092] In some embodiments, the vector is an “episomal expression vector” or “episode” capable of replicating within a host cell and persisting as an extrachromosomal segment of DNA within the host cell in the presence of appropriate selective pressure (see, e.g., Conese et al., Gene Therapy, 11: 1735–1742 (2004)). Representative commercially available episomal expression vectors include, among others, episomal plasmids utilizing Epstein-Barr nuclear antigen 1 (EBNA1) and the Epstein-Barr virus (EBV) origin of replication (oriP). The vectors pREP4, pCEP4, pREP7 and pcDNA3.1 from Invitrogen (Carlsbad, CA) and PB-CMV from Stratagene (La Jolla, CA) are non-restrictive examples of an episomal vector that uses the T antigen and the SV40 origin of replication instead of EBNAl and oriP.

[0093] In some embodiments, the vector is an “integrating expression vector” that can randomly integrate into host cell DNA or include a recombination site to enable recombination between the expression vector and a specific site in the host cell’s chromosomal DNA. Such integrating expression vectors can utilize the endogenous expression control sequences of the host cell’s chromosomes to induce the expression of the desired protein. Examples of vectors that integrate at a specific site include components of the flp-in system from Invitrogen (Carlsbad, CA) (e.g., pcDNA™5 / FRT) or the cre-LOX system, as found in the pExchange-6 core vectors from Stratagene (La Jolla, CA). Examples of vectors that randomly integrate into host cell chromosomes include pcDNA3.1 (when introduced in the absence of T-antigen) from Invitrogen (Carlsbad, CA) and PCI or pFNI OA (ACT) FLEXI™ from Promega (Madison, WI).

[0094] In some embodiments, the vector is a viral vector. Representative viral expression vectors include, among others, adenovirus-based vectors (e.g., the adenovirus-based per.C6 system available from Crucell, Inc. (Leiden, Netherlands)), lentivirus-based vectors (e.g., the lentivirus-based pLPl from Life Technologies (Carlsbad, CA)), and retroviral vectors (e.g., the pFB-ERV plus pCFB-EGagena (La) from Jollata, CA). In a preferred aspect, the viral vector is a lentivirus vector.

[0095] The vector comprising the nucleic acid according to the invention, which encodes the CAR, can be introduced into a host cell capable of expressing the encoded CAR, including any suitable prokaryotic or eukaryotic cell. Preferred host cells are those that can be easily and reliably cultured, exhibit relatively rapid growth rates, have well-characterized expression systems, and can be easily and efficiently transformed or transfected.

[0096] Within the scope of the present invention, the term "host cell" refers to any cell type that can contain the expression vector. The host cell can be a eukaryotic cell, e.g., plant, animal, fungi, or algae, or it can be a prokaryotic cell, e.g., bacteria or protozoa. The host cell can be a cultured cell or a primary cell, i.e., isolated directly from an organism, e.g., a human. The host cell can be an adherent cell or a suspended cell, i.e., a cell growing in suspension. Suitable host cells are known in the prior art and include, for example, DH5α E. coli cells, Chinese hamster ovary cells, monkey VERO cells, COS cells, HEK 293 cells, and the like. In a preferred aspect, the host cells are HEK 293 cells. In some embodiments, the HEK 293 cells are derived from the ATCC SD-3515 line.In some embodiments, the HEK 293 cells are derived from the IU-VPF-MCB line. In some embodiments, the HEK 293 cells are derived from the IU-VPF-MWCB line. In some embodiments, the host cell can be a peripheral blood lymphocyte (PBL), a peripheral blood mononuclear cell (PBMC), or a natural killer (NK) cell. Preferably, the host cell is an NK (natural killer) cell. Particularly preferably, the host cell is a T cell.

[0097] For amplification or replication of the recombinant expression vector, the host cell can be a prokaryotic cell, e.g., a DH5α cell. For the production of a virus from a viral expression vector, the host cell can be a eukaryotic cell, e.g., a HEK 293 cell. For the production of a recombinant CAR, the host cell can be a mammalian cell. Preferably, the host cell is a human cell. The host cell can be of any cell type, originate from any tissue type, and be at any developmental stage. Methods for selecting suitable mammalian host cells and methods for transforming, culturing, amplifying, screening, and purifying cells are known in the art.

[0098] In some embodiments, the disclosure provides an isolated host cell that expresses the nucleic acid sequence encoding the CAR described herein.

[0099] In some embodiments, the host cell is a T cell. The T cell of the disclosure can be any T cell, such as a cultured T cell, e.g., a primary T cell, or a T cell from a cultured T cell line, or a T cell obtained from a mammal. If obtained from a mammal, the T cell can be obtained from numerous sources, including, but not limited to, blood, bone marrow, lymph nodes, thymus, or other tissues or fluids. T cells can also be enriched or purified. The T cell is preferably a human T cell (e.g., isolated from a human). The T cell can be from any stage of development, including, but not limited to, a CD4+ / CD8+ double-positive T cell, a CD4+ helper T cell, etc. B. Th and Th2 cells, a CD8+ T cell (e.g. a cytotoxic T cell), a tumor-infiltrating cell, a memory T cell, a naive T cell and the like.In one aspect, the T cell is either a CD8+ T cell or a CD4+ T cell. T cell lines are available, for example, from the American Type Culture Collection (ATCC, Manassas, VA) and the German Collection of Microorganisms and Cell Cultures (DSMZ), and include, for example, Jurkat cells (ATCC TIB-152), Sup-Tl cells (ATCC CRL-1942), RPMI 8402 cells (DSMZ ACC-290), Karpas 45 cells (DSMZ ACC-545), and derivatives thereof.

[0100] In some embodiments, the host cell is a natural killer (NK) cell. NK cells are a type of cytotoxic lymphocyte that plays a role in the innate immune system. Defined as large granular lymphocytes, NK cells represent a third cell type, distinct from the common lymphoid precursor that also gives rise to B and T lymphocytes (see, e.g., Immunobiology, 5th ed., Janeway et al., eds., Garland Publishing, New York, NY (2001)). NK cells differentiate and mature in the bone marrow, lymph nodes, spleen, tonsils, and thymus. Upon maturation, NK cells enter the bloodstream as large lymphocytes containing characteristic cytotoxic granules. NK cells are able to recognize and kill some abnormal cells, such as some tumor cells and virus-infected cells, and are considered important for the innate immune defense against intracellular pathogens.As described above in relation to T cells, the NK cell can be any NK cell, such as a cultured NK cell (e.g., a primary NK cell), an NK cell derived from a cultured NK cell line, or an NK cell obtained from a mammal. If obtained from a mammal, the NK cell can be derived from numerous sources, including, but not limited to, blood, bone marrow, lymph nodes, thymus, or other tissues or fluids. NK cells can also be enriched or purified. The NK cell is preferably a human NK cell (e.g., isolated from a human). NK cell lines are available, for example, from the American Type Culture Collection (ATCC, Manassas, VA) and subsequently from other sources. B. NK-92 cells (ATCC CRL-2407), NK92MI cells (ATCC CRL-2408) and derivatives thereof.

[0101] In some embodiments, the nucleic acid sequences encoding a CAR can be introduced into a cell by "transfection," "transformation," or "transduction." "Transfection," "transformation," or "transduction," as used herein, refers to the introduction of one or more exogenous polynucleotides into a host cell using physical or chemical methods.

[0102] Many transfection techniques are known in the art and include, for example, calcium phosphate DNA precipitation (see, e.g., Murray EJ (ed.), Methods in Molecular Biology, Vol. 7, Gene Transfer and Expression Protocols, Humana Press (1991)); DEAE-dextran; electroporation; cationic liposome-mediated transfection; tungsten particle-facilitated microparticle bombardment (Johnston, Nature, 346: 776-777 (1990)); and strontium phosphate DNA precipitation (Brash et al., Mol. Cell Biol., 7: 2031-2034 (1987)). Phage or virus vectors can be introduced into host cells after the growth of infectious particles into suitable packaging cells, many of which are commercially available. Chimeric antigen receptors

[0103] International patent publication no. WO 2018 / 028647 is incorporated herein in its entirety by reference. US patent publication no. 2018 / 0230225 is incorporated herein in its entirety by reference. Both publications describe chimeric antigen receptors (CARs) targeting BCMA that are suitable in the present disclosure.

[0104] The disclosure provides methods for treating a subject with cells expressing a chimeric antigen receptor (CAR). The CAR comprises an extracellular antigen-binding domain containing one or more single-domain antibodies. In various aspects and embodiments, a CAR targeting BCMA (here also referred to as the "BCMA-CAR") is provided, comprising a polypeptide that includes: (a) an extracellular antigen-binding domain containing an anti-BCMA binding unit; (b) a transmembrane domain; and (c) an intracellular signaling domain. In some embodiments, the anti-BCMA binding unit is camelid, chimeric, human, or humanized. In some embodiments, the intracellular signaling domain comprises a primary intracellular signaling domain of an immune effector cell (such as a T cell). In some embodiments, the primary intracellular signaling domain is derived from CD4.In some embodiments, the primary intracellular signaling domain is derived from CD3-zeta. In some embodiments, the intracellular signaling domain includes a costimulatory signaling domain. In some embodiments, the costimulatory signaling domain is derived from a costimulatory molecule selected from the group consisting of CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, ligands of CD83, and combinations thereof. In certain embodiments, the transmembrane domain is derived from CD137.

[0105] In some embodiments, the BCMA-CAR further comprises a joint domain (such as a CD8-alpha joint domain) located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the BCMA-CAR further comprises a signal peptide (such as a CD8-alpha signal peptide) located at the N-terminus of the polypeptide. In some embodiments, the polypeptide, from the N-terminus to the C-terminus, comprises: a CD8-alpha signal peptide, the extracellular antigen-binding domain, a CD8-alpha joint domain, a CD28-derived first costimulatory signaling domain, a CD137-derived second costimulatory signaling domain, and a CD4-derived primary intracellular signaling domain.In some embodiments, the polypeptide comprises, from the N-terminus to the C-terminus: a CD8-alpha signaling peptide, the extracellular antigen-binding domain, a CD8-alpha joint domain, a CD8-alpha transmembrane domain, a second costimulatory signaling domain derived from CD137, and a primary intracellular signaling domain derived from CD3-zeta. In some embodiments, the BCMA-CAR is monospecific. In some embodiments, the BCMA-CAR is monovalent.

[0106] The present application also provides CARs comprising two or more (including, inter alia, one of 2, 3, 4, 5, 6, or more) binding units that bind specifically to an antigen, such as BCMA. In some embodiments, one or more of the binding units are antigen-binding fragments. In some embodiments, one or more of the binding units comprise single-domain antibodies. In some embodiments, one or more of the binding units comprise a VHH.

[0107] In some embodiments, the CAR is a multivalent (such as bivalent, trivalent or a higher number of valences) CAR comprising a polypeptide that includes: (a) an extracellular antigen-binding domain containing multiple (such as at least about one of 2, 3, 4, 5, 6, or more) binding units that bind specifically to an antigen (such as a tumor antigen); (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0108] In some embodiments, the bonding units, such as VHHs (including multiple VHHs or the first VHH and / or the second VHH), are camelid, chimeric, human, or humanized. In some embodiments, the bonding units or VHHs are linked to each other via peptide bonds or peptide linkers. In some embodiments, each peptide linker is no more than about 50 amino acids long (such as no more than about one of 35, 25, 20, 15, 10, or 5).

[0109] In some embodiments, the first BCMA binding unit and / or the second BCMA binding unit is an anti-BCMA VHH. In some embodiments, the first BCMA binding unit is a first anti-BCMA VHH and the second BCMA binding unit is a second anti-BCMA VHH.

[0110] In some embodiments, the first BCMA binding unit and the second BCMA binding unit are linked together via a peptide linker. In some embodiments, the peptide linker comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the peptide linker comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 11.

[0111] In some embodiments, the CAR further comprises a hinge domain (such as a CD8-alpha hinge domain) located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the CAR further comprises a signal peptide (such as a CD8-alpha signal peptide) located at the N-terminus of the polypeptide.

[0112] Without being bound to any specific theory, CARs that are multivalent, or those that include an extracellular antigen-binding domain comprising a first BCMA-binding unit and a second BCMA-binding unit, may be particularly well-suited to target multimeric antigens via synergistic binding through the different antigen-binding sites, or to increase the binding affinity or avidity to the antigen. Improved avidity can allow for a substantial reduction in the dose of CAR-T cells required to achieve a therapeutic effect, such as a dose in the range of 4.0 × 10⁻⁶. 4 up to 1.0 × 10 6 CAR-T cells per kilogram of subject mass or 3.0 × 10 6 up to 1.0 × 10 8CAR-T expressing total cells. Monovalent CARs, such as bb2121, may need to be dosed at 5 to 10 times these amounts to achieve a comparable effect. In various formulations, reduced dosage ranges can substantially reduce cytokine release syndrome (CRS) and other potentially dangerous side effects of CAR-T therapy.

[0113] The various binding units (e.g., an extracellular antigen-binding domain comprising a first BCMA binding unit and a second BCMA binding unit) in the CARs described herein can be linked to one another via peptide linkers. The peptide linkers connecting different binding units (such as VHHs) can be the same or different. Different domains of the CARs can also be linked to one another via peptide linkers. In some embodiments, the binding units (such as VHHs) are directly linked to one another without peptide linkers.

[0114] The peptide linker in the CAR described herein can have any suitable length. In some embodiments, the peptide linker is at least approximately one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 75, 100 or more amino acids long. In some embodiments, the peptide linker is no more than about one of 100, 75, 50, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5 or less amino acids long.In some embodiments, the length of the peptide linker is any length from about 1 amino acid to about 10 amino acids, about 1 amino acid to about 20 amino acids, about 1 amino acid to about 30 amino acids, about 5 amino acids to about 15 amino acids, about 10 amino acids to about 25 amino acids, about 5 amino acids to about 30 amino acids, about 10 amino acids to about 30 amino acids, about 30 amino acids to about 50 amino acids, about 50 amino acids to about 100 amino acids, or about 1 amino acid to about 100 amino acids.

[0115] The CARs of the present application comprise a transmembrane domain which may be directly or indirectly linked to the extracellular antigen-binding domain.

[0116] The CAR can include a T-cell activation unit. The T-cell activation unit can be any suitable unit derived from or obtained from any suitable molecule. In one aspect, the T-cell activation unit includes, for example, a transmembrane domain. The transmembrane domain can be any transmembrane domain derived from or obtained from any molecule known in the art. For example, the transmembrane domain can be obtained from or derived from a CD8α molecule or a CD28 molecule. Without being bound to any theory, CD8 is a transmembrane glycoprotein that serves as a coreceptor for the T-cell receptor (TCR) and is mainly expressed on the surface of cytotoxic T cells. The most common form of CD8 exists as a dimer consisting of a CD8α and a CD8β chain.CD28 is expressed on T cells and provides co-stimulatory signals required for T-cell activation. CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2). In a preferred aspect, CD8α and CD28 are human.

[0117] In addition to the transmembrane domain, the T-cell activation unit may further comprise an intracellular (i.e., cytoplasmic) T-cell signaling domain. The intracellular T-cell signaling domain may be obtained from or derived from a CD28 molecule, a CD3 zeta (FcRγ) molecule or modified versions thereof, a human FC receptor gamma (ζ) chain, a CD27 molecule, an OX40 molecule, a 4-1BB molecule, or other known prior art intracellular signaling molecules. Without being bound to any theory, it is true that: (1) CD28 is a T-cell marker important for T-cell co-stimulation; (2) CD3ζ associates with TCRs to generate a signal and contains immunoreceptor tyrosine-based activation motifs (ITAMs). and (3) 4-1BB, also known as CD137, transmits a strong costimulatory signal to T cells, thereby promoting differentiation and improving the long-term survival of T lymphocytes.In one preferred aspect, CD28, CD3 zeta, 4-1BB, OX40, and CD27 are human.

[0118] The T-cell activation domain of a CAR encoded by the nucleic acid sequences disclosed herein can comprise any of the transmembrane domains mentioned above and any combination of any of the intercellular T-cell signaling domains mentioned above. For example, the nucleic acid sequences disclosed herein can encode a CAR comprising a CD28 transmembrane domain and intracellular T-cell signaling domains of CD28 and CD3 zeta. Alternatively, the nucleic acid sequences disclosed herein can, for example, encode a CAR comprising a CD8α transmembrane domain and intracellular T-cell signaling domains of CD28, CD3 zeta, the FC receptor gamma (FcRγ) chain, and / or 4-1BB.

[0119] In some embodiments, the CAR polypeptide further comprises a signal peptide located at the N-terminus of the polypeptide. In some embodiments, the signal peptide is derived from CD8-alpha. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the signal peptide comprises a polypeptide defined by the nucleic acid sequence of SEQ ID NO: 9.

[0120] In certain embodiments, the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 6. In certain embodiments, the transmembrane domain comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 14.

[0121] In some embodiments, the intracellular signaling domain comprises a primary intracellular signaling domain of an immune effector cell. In some embodiments, the intracellular signaling domain is derived from CD3ζ. In some embodiments, the intracellular signaling domain comprises at least one costimulatory signaling domain. In some embodiments, the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the intracellular signaling domain comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 16. In some embodiments, the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 7. In some embodiments, the intracellular signaling domain comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 15.

[0122] In some embodiments, the CAR polypeptide further comprises a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the hinge domain comprises the amino acid sequence of SEQ ID NO: 5. In some embodiments, the hinge domain comprises a polypeptide encoded by the nucleic acid sequence of SEQ ID NO: 13.

[0123] In some embodiments, the CAR comprises one, more, or all of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8. In one aspect, the CAR comprises SEQ ID NO: 17. In some embodiments, the CAR comprises a polypeptide encoded by the nucleic acid sequence of one, more, or all of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16.

[0124] In preferred embodiments, the CAR comprises a first VHH domain comprising CDR1, CDR2, and CDR3 of the VHH domain, comprising the amino acid sequence of SEQ ID NO: 2, and a second VHH domain comprising CDR1, CDR2, and CDR3 of the VHH domain, comprising the amino acid sequence of SEQ ID NO: 4. In preferred embodiments, the first VHH domain is linked via a linker comprising the amino acid sequence of SEQ ID NO: 3. In particularly preferred embodiments, the first VHH domain comprises CDR1 comprising the amino acid sequence of SEQ ID NO: 18, CDR2 comprising the amino acid sequence of SEQ ID NO: 19, and CDR3 comprising the amino acid sequence of SEQ ID NO: 20, and the second VHH domain comprises CDR1 comprising the amino acid sequence of SEQ ID NO: 21, CDR2 comprising the amino acid sequence of SEQ ID NO: 22, and CDR3 comprising the Amino acid sequence of SEQ ID NO: 23.In further preferred embodiments, the CAR comprises a first VHH domain comprising the amino acid sequence of SEQ ID NO: 2, and a second VHH domain comprising the amino acid sequence of SEQ ID NO: 4. Immune effector cell compositions

[0125] "Immune effector cells" are immune cells capable of performing immune effector functions. In some embodiments, the immune effector cells express at least FcγRIII and perform an ADCC effector function. Examples of immune effector cells that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, neutrophils, and eosinophils. In some embodiments, the immune effector cells are T cells. In some embodiments, the T cells are autologous T cells. In some embodiments, the T cells are allogeneic T cells. In some embodiments, the T cells are CD4+ / CD8-, CD4- / CD8+, CD4+ / CD8+, CD4- / CD8-, or combinations thereof. In some embodiments, the T cells produce IL-2, TFN and / or TNF after expression of the CAR and binding to the target cells, such as CD20+ or ​​CD19+ tumor cells.In some embodiments, the CD8+ T cells lyse antigen-specific target cells after expression of the CAR and binding to the target cells.

[0126] Biological methods for introducing the vector into an immune effector cell include the use of DNA and RNA vectors. Viral vectors have become the most widely used method for inserting genes into mammalian cells, such as human cells. Chemical means for introducing the vector into an immune effector cell include colloidal dispersion systems, such as macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems, including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An example of a colloidal system for use as an in vitro delivery vehicle is a liposome (e.g., an artificial membrane vesicle).

[0127] This provides dosage forms that are 3.0 × 10 7 up to 1.0 × 10 8CAR-T cells comprise a CAR comprising a polypeptide that includes: (a) an extracellular antigen-binding domain comprising a first BCMA-binding unit that binds specifically to a first epitope of BCMA and a second BCMA-binding unit that binds specifically to a second epitope of BCMA; (b) a transmembrane domain; and (c) an intracellular signaling domain, wherein the first and second epitopes are different. In certain embodiments, the dosage form comprises 3.0 × 10 7 up to 4.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 3.5 × 10 7 up to 4.5 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 4.0 × 10 7 up to 5.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 4.5 × 10 7 up to 5.5 × 10 7of the CAR-T cells. In certain embodiments, the dosage form comprises 5.0 × 10 7 up to 6.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 5.5 × 10 7 up to 6.5 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 6.0 × 10 7 up to 7.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 6.5 × 10 7 up to 7.5 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 7.0 × 10 7 up to 8.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 7.5 × 10 7 up to 8.5 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 8.0 × 10 7 up to 9.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 8.5 × 10 7 up to 9.5 × 10 7of the CAR-T cells. In certain embodiments, the dosage form comprises 9.0 × 10 7 up to 1.0 × 10 8 the CAR-T cells.

[0128] In some embodiments, dosage forms are provided that contain 3.0 × 10 7 up to 1.0 × 10 8 The engineered immune effector cells (such as T cells) comprise a CAR comprising a polypeptide, comprising: (a) an extracellular antigen-binding domain comprising a first anti-BCMA VHH that binds specifically to a first epitope of BCMA and a second anti-BCMA VHH that binds specifically to a second epitope of BCMA; (b) a transmembrane domain; and (c) an intracellular signaling domain, wherein the first and second epitopes are different. In certain embodiments, the dosage form comprises 3.0 × 10 7 up to 4.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 3.5 × 10 7 up to 4.5 × 107 of the CAR-T cells. In certain embodiments, the dosage form comprises 4.0 × 10 7 up to 5.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 4.5 × 10 7 up to 5.5 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 5.0 × 10 7 up to 6.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 5.5 × 10 7 up to 6.5 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 6.0 × 10 7 up to 7.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 6.5 × 10 7 up to 7.5 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 7.0 × 10 7 up to 8.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 7.5 × 10 7 up to 8.5 × 107 of the CAR-T cells. In certain embodiments, the dosage form comprises 8.0 × 10 7 up to 9.0 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 8.5 × 10 7 up to 9.5 × 10 7 of the CAR-T cells. In certain embodiments, the dosage form comprises 9.0 × 10 7 up to 1.0 × 10 8 the CAR-T cells.

[0129] In some embodiments, the cell population of the CAR-T cell dosing formulations described here comprises a T cell or a population of T cells, e.g., at various stages of differentiation. Stages of T-cell differentiation include naive T cells, central stem memory T cells, central storage T cells, effector storage T cells, and terminal effector T cells, ranging from least to most differentiated. Following antigen exposure, naive T cells proliferate and differentiate into memory T cells, e.g., central stem memory T cells and central storage T cells, which then differentiate into effector storage T cells. Upon receiving appropriate T-cell receptor, costimulatory, and inflammatory signals, memory T cells further differentiate into terminal effector T cells. See, e.g., Restifo. Blood. 124.4(2014):476-77; and Joshi et al. J. Immunol. 180.3(2008):1309-15.

[0130] Naive T cells can exhibit the following cell surface marker expression pattern: CCR7+, CD62L+, CD45RO-, CD95-. Central stem memory (Tscm) T cells can exhibit the following cell surface marker expression pattern: CCR7+, CD62L+, CD45RO-, CD95+. Central storage (TCM) T cells can exhibit the following cell surface marker expression pattern: CCR7+, CD62L+, CD45RO+, CD95+. Effector storage (Tem) T cells can exhibit the following cell surface marker expression pattern: CCR7-, CD62L-, CD45RO+, CD95+. Terminal effector (Teff) T cells can exhibit the following cell surface marker expression pattern: CCR7-, CD62L-, CD45RO-, CD95+. See, e.g., Gattinoni et al., Nat. Med. 17(2011):1290-7; and Flynn et al Clin. Translat. Immunol. 3(2014):e20. Pharmaceutical compositions and formulations

[0131] Furthermore, the present application provides pharmaceutical compositions comprising any of the anti-BCMA antibodies disclosed or any of the engineered immune effector cells, any of the CARs described herein (such as BCMA-CARs), and a pharmaceutically acceptable carrier. Pharmaceutical compositions can be prepared by mixing any of the immune effector cells described herein, of the desired purity, with optionally pharmaceutically acceptable carriers, excipients, or stabilizers (Remington's Pharmaceutical Sciences 16th Edition, OSOL, A. Ed (1980)) in the form of lyophilized formulations or aqueous solutions. In certain embodiments, a pharmaceutical composition of CAR-T cells further comprises an excipient selected from dimethyl sulfoxide (DMSO) or Dextran-40. In some embodiments, the formulation provided herein comprises 5% DMSO.

[0132] The compositions described herein can be administered as part of a pharmaceutical composition comprising one or more carriers. The choice of carrier is determined in part by the specific nucleic acid sequence, vector, or host cells expressing the CARs disclosed herein, as well as by the specific method used to administer the nucleic acid sequence, vector, or host cells expressing the CARs disclosed herein. Accordingly, there are a variety of suitable formulations of the pharmaceutical compositions disclosed herein.

[0133] For example, pharmaceutical compositions may contain preservatives. Suitable preservatives may include, for example, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. A mixture of two or more preservatives may be used if necessary. The preservative, or mixtures thereof, is typically present in an amount of approximately 0.0001 to approximately 2% by weight of the total composition.

[0134] Furthermore, buffering agents may be used in the formulations. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. A mixture of two or more buffering agents may be used if necessary. The buffering agent or mixtures thereof are typically present in an amount of approximately 0.001 to approximately 4% by weight of the total formulation.

[0135] The compositions comprising the nucleic acid sequence encoding the CARs disclosed herein, or host cells expressing the CARs disclosed herein, can be formulated as an inclusion complex, such as a cyclodextrin inclusion complex, or as a liposome. Liposomes can be used to direct the host cells (e.g., T cells or NK cells) or the nucleic acid sequences disclosed herein to a specific tissue. Liposomes can also be used to increase the half-life of the nucleic acid sequences disclosed herein. Many methods are available for the production of liposomes, such as those described in Szoka et al., Ann. Rev. Biophys. Bioeng., 9: 467 (1980) and US patents 4,235,871; 4,501,728; 4,837,028 and 5,019,369.The compositions can employ time-release, delayed-release, and delayed-release delivery systems, such that the delivery of the compositions disclosed herein occurs before and with sufficient time to sensitize the treatment site. Many types of release delivery systems are available and known to the scientific community. Such systems can avoid repeated administrations of the composition, thereby increasing convenience for both the patient and the physician, and may be particularly suitable for certain compositional aspects and embodiments of the disclosure.

[0136] In certain embodiments, the CAR-T cells are formulated in a dose of approximately 1.0 × 10 5 up to 2.0 × 10 5 cells / kg, 1.5 × 10 5 up to 2.5 × 10 5 cells / kg, 2.0 × 10 5 up to 3.0 × 10 5 cells / kg, 2.5 × 10 5 up to 3.5 × 10 5cells / kg, 3.0 × 10 5 up to 4.0 × 10 5 cells / kg, 3.5 × 10 5 up to 4.5 × 10 5 cells / kg, 4.0 × 10 5 up to 5.0 × 10 5 cells / kg, 4.5 × 10 5 up to 5.5 × 10 5 cells / kg, 5.0 × 10 5 up to 6.0 × 10 5 cells / kg, 5.5 × 10 5 up to 6.5 × 10 5 cells / kg, 6.0 × 10 5 up to 7.0 × 10 5 cells / kg, 6.5 × 10 5 up to 7.5 × 10 5 cells / kg, 7.0 × 10 5 up to 8.0 × 10 5 cells / kg, 7.5 × 10 5 up to 8.5 × 10 5 cells / kg, 8.0 × 10 5 up to 9.0 × 10 5 cells / kg, 8.5 × 10 5 up to 9.5 × 10 5 cells / kg, 9.0 × 10 5 up to 1.0 × 10 6 cells / kg. In one preferred aspect, the dose is approximately 0.75 x 10 6 -cells / kg formulated. In certain embodiments, the CAR-T cells are formulated at a dose of less than 1.0 × 10 8-cells per subject. Preferably, the dose is administered as a single infusion. Treatment methods and applications

[0137] The present application also relates to methods and compositions for use in cell immunotherapy.

[0138] In some aspects, a method for treating cancer in a subject with multiple myeloma who has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD, is provided herein. In some embodiments, the subject has received one prior line of therapy. In some embodiments, the subject has received two prior lines of therapy. In some embodiments, the subject has received three prior lines of therapy.

[0139] In preferred embodiments, the subject has received prior treatment with an IMiD as part of one or more of the 1 to 3 prior lines of therapy. In some embodiments, the IMiD is lenalidomide. In preferred embodiments, the patient is lenalidomide-refractory. In some embodiments, the prior treatment includes pomalidomide. In some embodiments, the prior IMiD line of therapy includes a combination of lenalidomide and pomalidomide. In some embodiments, the subject has received prior treatment with a proteasome inhibitor as part of one or more of the 1 to 3 prior lines of therapy. In some embodiments, the proteasome inhibitor is bortezomib, carfilzomib, ixazomib, or any combination thereof. In some embodiments, the subject has received prior treatment with an anti-CD38 antibody as part of one or more of the 1 to 3 prior lines of therapy.In some embodiments, the anti-CD38 antibody is daratumumab and / or isatuximab. In some embodiments, prior treatment includes an IMiD (e.g., lenalidomide), a proteasome inhibitor, and an anti-CD38 antibody (i.e., three prior lines of therapy). In some embodiments, the subject has received one prior line of therapy, including lenalidomide, and is lenalidomide-refractory, and may have received one or two additional lines of therapy. In some embodiments, the subject has received at least one prior line of therapy, including lenalidomide and a proteasome inhibitor, and may have received one or two additional lines of therapy.

[0140] The therapy may be used, if appropriate, to treat the subject who has a high-risk feature, including, for example, a cytogenetic abnormality, International Staging System (ISS) Stage III, and / or soft tissue plasmacytomas. In some embodiments, the high-risk feature is a cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality. In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from a group that includes Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic abnormality includes Gain / amp(1q). In some embodiments, the cytogenetic abnormality includes del(17p). In some aspects, the cytogenetic abnormality includes t(4;14).In some embodiments, the cytogenetic abnormality comprises t(14;16). t(4;14) and t(14;16) are translocations in which portions of the chromosome are exchanged. Del(17p) is a loss of part of the short arm of chromosome 17. Gain / amp(1q) indicates a gain (e.g., 3 total copies) or amplification (e.g., > 3 total copies) of a portion of the long arm of chromosome 1. In some embodiments, the subject has at least two cytogenetic abnormalities. In other embodiments, the subject has two, three, four, five, or more cytogenetic abnormalities. In other embodiments, the cytogenetic abnormality is a standard-risk cytogenetic abnormality. In some embodiments, the high-risk feature is International Staging System (ISS) Level III. In some embodiments, the high-risk feature is soft tissue plasmacytomas.

[0141] In other aspects, the procedure provided here involves first determining whether the subject has a high-risk feature, wherein the high-risk feature is a cytogenetic abnormality, International Staging System (ISS) stage III, and / or soft tissue plasmacytomas; and then administering the compositions provided here to the subject determined to have the high-risk feature. In some aspects, the subject has multiple myeloma, has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD. In some embodiments, the IMiD is lenalidomide. In some embodiments, the high-risk feature is a cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality.In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from a group that includes Gain / AMP(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic abnormality includes Gain / AMP(1q). In some embodiments, the cytogenetic abnormality includes del(17p). In some embodiments, the cytogenetic abnormality includes t(4;14). In some embodiments, the cytogenetic abnormality includes t(14;16). In some embodiments, the subject has at least two cytogenetic abnormalities. In other embodiments, the subject has two, three, four, five, or more cytogenetic abnormalities. In other embodiments, the cytogenetic abnormality is a standard-risk cytogenetic abnormality.In some embodiments, the high-risk feature is International Staging System (ISS) Stage III. In some embodiments, the high-risk feature is soft tissue plasmacytomas. In some embodiments, the subject has received one prior line of therapy. In some embodiments, the subject has received two prior lines of therapy. In some embodiments, the subject has received three prior lines of therapy. In some embodiments, the prior treatment includes pomalidomide. In some aspects, the prior treatment further includes a proteasome inhibitor, wherein the proteasome inhibitor is optionally bortezomib, carfilzomib, ixazomib, or any combination thereof. In some embodiments, the prior treatment further includes an anti-CD38 antibody, wherein the anti-CD38 antibody is optionally daratumumab and / or isatuximab. In some embodiments, the prior treatment includes an IMiD (e.g.,lenalidomide), a proteasome inhibitor and an anti-CD38 antibody.

[0142] In other aspects, a method for the selective treatment of a subject with the compositions provided herein is provided, comprising administering the composition to the subject who has been determined to have a high-risk feature such as a cytogenetic abnormality, International Staging System (ISS) stage III, and / or soft tissue plasmacytomas. In some embodiments, the composition provided herein is for use in the treatment of a subject who has been determined to have a high-risk feature such as a cytogenetic abnormality, International Staging System (ISS) stage III, and / or soft tissue plasmacytomas. In some embodiments, the subject has multiple myeloma, has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD. In some embodiments, the IMiD is lenalidomide.In some embodiments, the high-risk feature is a cytogenetic anomaly. In some embodiments, the cytogenetic anomaly is a high-risk cytogenetic anomaly. In some embodiments, the subject has one or more high-risk cytogenetic anomalies selected from a group that includes Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof. In some embodiments, the cytogenetic anomaly includes Gain / amp(1q). In some embodiments, the cytogenetic anomaly includes del(17p). In some embodiments, the cytogenetic anomaly includes t(4;14). In some embodiments, the cytogenetic anomaly includes t(14;16). In some embodiments, the subject has at least two cytogenetic anomalies. In other embodiments, the subject exhibits two, three, four, five or more cytogenetic abnormalities.In other embodiments, the cytogenetic abnormality is a standard-risk cytogenetic abnormality. In some embodiments, the high-risk feature is International Staging System (ISS) Grade III. In some embodiments, the high-risk feature is soft tissue plasmacytomas. In some embodiments, the subject has received one prior line of therapy. In some embodiments, the subject has received two prior lines of therapy. In some embodiments, the subject has received three prior lines of therapy. In some embodiments, the prior treatment includes pomalidomide. In some embodiments, the prior treatment further includes a proteasome inhibitor, the proteasome inhibitor being optionally bortezomib, carfilzomib, ixazomib, or any combination thereof.In some embodiments, the prior treatment further comprises an anti-CD38 antibody, optionally being daratumumab and / or isatuximab. In some embodiments, the prior treatment comprises an IMiD (e.g., lenalidomide), a proteasome inhibitor, and an anti-CD38 antibody.

[0143] In some embodiments of the various methods or uses provided herein, the subject further receives bridge therapy, and the bridge therapy may optionally be selected by the physician. The bridge therapy may include pomalidomide, bortezomib, dexamethasone, daratumumab, or any combination thereof. It may include pomalidomide, bortezomib, and dexamethasone. Another exemplary bridge therapy includes daratumumab, pomalidomide, and dexamethasone. In some embodiments, the subject receives the bridge therapy from approximately every 20 days to approximately every 30 days, for example, approximately every 21 days or approximately every 28 days. In some embodiments, the subject receives at least one, two, three, four, or more bridge therapies.

[0144] In certain embodiments, the bridge therapy comprises a 28-day cycle consisting of daratumumab on days 1, 8, 15, and 22; pomalidomide for 21 days; and dexamethasone on days 1, 8, 15, and 22. For example, the bridge therapy may include 1800 mg of daratumumab on days 1, 8, 15, and 22; 4 mg / day of pomalidomide for 21 days; and 40 mg of dexamethasone on days 1, 8, 15, and 22. In these embodiments, the daratumumab may be administered subcutaneously, the pomalidomide orally, and the dexamethasone orally or intravenously.

[0145] In other embodiments, the bridge therapy comprises a 21-day cycle including bortezomib on days 1, 4, 8, and 11, pomalidomide for 14 days, and dexamethasone on days 1, 2, 4, 5, 8, 9, 11, and 12. For example, the bridge therapy may consist of 1.3 mg / m² 2The regimen includes bortezomib on days 1, 4, 8, and 11; 4 mg / day pomalidomide for 14 days; and 20 mg dexamethasone on bridge days 1, 2, 4, 5, 8, 9, 11, and 12. In these embodiments, the bortezomib can be administered subcutaneously, the pomalidomide can be administered orally, and the dexamethasone can be administered orally.

[0146] In some embodiments, the subject has further received lymphodepletion therapy, for example, following a bridging therapy disclosed herein. In some embodiments, the lymphodepletion therapy comprises daily cyclophosphamide and / or fludarabine. In one aspect, the lymphodepletion therapy comprises daily cyclophosphamide and fludarabine. In some embodiments, the lymphodepletion therapy comprises cyclophosphamide at a concentration of approximately 300 mg / m³. 2 and fludarabine at a concentration of approximately 30 mg / m³ 2 daily for 3 days.

[0147] In some embodiments of the various methods or uses provided herein, the dose of the CAR-T cells is 0.5-1.0 × 10 6 -cells / kg body weight of the subject. In a preferred aspect, the dose of CAR-T cells is approximately 0.75 × 10 6 -cells / kg body weight of the subject. In some embodiments, the method includes administering the dose of CAR-T cells approximately 5 to 7 days after the start of lymphodepletion therapy. Preferably, the dose is administered as a single infusion. In some embodiments, a single intravenous infusion of CAR-T cells of 0.75 × 10⁻⁵ is administered 5–7 days after the start of lymphodepletion. 6 -cells / kg administered.

[0148] Any of the anti-BCMA VHHs, CARs, and engineered immune effector cells (such as CAR-T cells) described herein can be used in the procedure to treat cancer. In some embodiments, the immune effector cells are autologous. In some embodiments, the immune effector cells are allogeneic.

[0149] In certain embodiments, the CAR-T cells are administered in a dose of approximately 1.0 × 10 5 up to 2.0 × 10 5 cells / kg, 1.5 × 10 5 up to 2.5 × 10 5 cells / kg, 2.0 × 10 5 up to 3.0 × 10 5 cells / kg, 2.5 × 10 5 up to 3.5 × 10 5 cells / kg, 3.0 × 10 5 up to 4.0 × 10 5 cells / kg, 3.5 × 10 5 up to 4.5 × 10 5 cells / kg, 4.0 × 10 5 up to 5.0 × 10 5 cells / kg, 4.5 × 10 5 up to 5.5 × 10 5 cells / kg, 5.0 × 10 5 up to 6.0 × 10 5 cells / kg, 5.5 × 10 5up to 6.5 × 10 5 cells / kg, 6.0 × 10 5 up to 7.0 × 10 5 cells / kg, 6.5 × 10 5 up to 7.5 × 10 5 cells / kg, 7.0 × 10 5 up to 8.0 × 10 5 cells / kg, 7.5 × 10 5 up to 8.5 × 10 5 cells / kg, 8.0 × 10 5 up to 9.0 × 10 5 cells / kg, 8.5 × 10 5 up to 9.5 × 10 5 cells / kg, 9.0 × 10 5 up to 1.0 × 10 5 cells / kg, 1.0 × 10 6 up to 2.0 × 10 6 cells / kg, 1.5 × 10 6 up to 2.5 × 10 6 cells / kg, 2.0 × 10 6 up to 3.0 × 10 6 cells / kg, 2.5 × 10 6 up to 3.5 × 10 6 cells / kg, 3.0 × 10 6 up to 4.0 × 10 6 cells / kg, 3.5 × 10 6 up to 4.5 × 10 6 cells / kg, 4.0 × 10 6 up to 5.0 × 10 6 cells / kg, 4.5 × 10 6 up to 5.5 × 10 6 cells / kg or 5.0 × 10 6 up to 6.0 × 10 6cells / kg. In a preferred aspect, the dose comprises approximately 0.75 × 10 6 -cells / kg. In certain embodiments, the CAR-T cells are administered at a dose of approximately 1.0 × 10 8 -cells per subject. Preferably, the dose is administered as a single infusion.

[0150] In certain embodiments, the CAR-T cells are administered at a dose of less than 1.0 × 10 8 Cells per subject. In certain embodiments, the CAR-T cells are administered at a dose of approximately 3.0 to 4.0 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 3.5 to 4.5 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 4.0 to 5.0 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 4.5 to 5.5 × 10⁻⁶. 7cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 5.0 to 6.0 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 5.5 to 6.5 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 6.0 to 7.0 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 6.5 to 7.5 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 7.0 to 8.0 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 7.5 to 8.5 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 8.0 to 9.0 × 10 7cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 8.5 to 9.5 × 10 7 cells. In certain embodiments, the CAR-T cells are administered at a dose of approximately 9.0 × 10 7 up to 1.0 × 10 8 administered to cells.

[0151] In certain embodiments, the CAR-T cells are administered at a dose of approximately 0.693 × 10 6 CAR-positive viable T cells / kg are administered. In certain formulations, the CAR T cells are administered at a dose of approximately 0.52 × 10 6 CAR-positive viable T cells / kg are administered. In certain formulations, the CAR T cells are administered at a dose of approximately 0.94 × 10 6 CAR-positive viable T cells / kg are administered. In certain formulations, the CAR T cells are administered at a dose of approximately 0.709 × 10 6CAR-positive viable T cells / kg are administered. In certain formulations, the CAR T cells are administered at a dose of approximately 0.51 × 10 6 CAR-positive viable T cells / kg are administered. In certain formulations, the CAR T cells are administered at a dose of approximately 0.95 × 10 6 CAR-positive viable T cells / kg are administered. In certain formulations, the CAR T cells are administered in an outpatient setting.

[0152] In some embodiments, the composition comprising CAR-T cells administered to the subject further includes an excipient selected from dimethyl sulfoxide (DMSO) or dextran-40. In some embodiments, the composition comprises 5% DMSO.

[0153] In certain embodiments, the CAR-T cells (e.g., in one of the preceding doses) are administered in one or more intravenous infusions. In certain embodiments, the CAR-T cells are administered via a single intravenous infusion. In certain embodiments, the single intravenous infusion is administered using a single bag of CAR-T cells. In certain embodiments, the administration of the single bag of CAR-T cells is completed between the time the single bag of CAR-T cells is thawed and three hours after the single bag of CAR-T cells has thawed. In certain embodiments, a single intravenous administration is performed using two bags of CAR-T cells.In certain embodiments, the administration of each of the two CAR-T cell bags is completed between the time when the first of the two CAR-T cell bags is thawed and three hours after the first bag of CAR-T cells has been thawed.

[0154] In certain embodiments, the time from initial apheresis to CAR-T cell administration is less than 41, 47, 54, 61, 68, 75, 82, 89, 96, 103, 110, 117, 124, 131, 138, 145, 152, 159, 166, or 167 days. In certain embodiments, the time from initial apheresis to CAR-T cell administration is greater than 41, 47, 54, 61, 68, 75, 82, 89, 96, 103, 110, 117, 124, 131, 138, 145, 152, 159, 166, or 167 days.

[0155] In certain embodiments, a lymphodepleting regimen precedes the administration of CAR-T cells. In certain embodiments, the lymphodepleting regimen includes the administration of cyclophosphamide and / or fludarabine. In certain embodiments, the lymphodepleting regimen is administered intravenously. In certain embodiments, the lymphodepleting regimen precedes the administration of CAR-T cells by 5 to 7 days. In certain embodiments, the lymphodepleting regimen precedes the administration of CAR-T cells by 2 to 4 days. According to certain embodiments, the lymphodepleting regimen includes the intravenous administration of cyclophosphamide and fludarabine 5 to 7 days prior to the administration of CAR-T cells. According to certain embodiments, the lymphodepleting regimen includes the intravenous administration of cyclophosphamide and fludarabine 2 to 4 days prior to the administration of CAR-T cells.In certain embodiments, the lymphodepleting regimen includes cyclophosphamide administered intravenously at 300 mg / m². 2 is administered. In certain formulations, the lymphodepleting regimen includes fludarabine, administered intravenously at 30 mg / m². 2 The lymphodepleting regimen is administered. In some formulations, it is performed daily for 3 days. In situations where CAR-T cell administration is delayed by more than 14 days, the lymphodepleting regimen can be repeated.

[0156] In certain embodiments, the CAR-T cell treatment method further includes treating the subject for cytokine release syndrome (CRS) within 3 days of CAR-T cell administration, without significantly reducing CAR-T cell expansion in vivo. In certain embodiments, CRS treatment includes administering an IL-6R inhibitor to the subject. In certain embodiments, the IL-6R inhibitor is an antibody. In certain embodiments, the IL-6 inhibitor inhibits IL-6R by binding to its extracellular domain. In certain embodiments, the IL-6R inhibitor prevents IL-6 from binding to IL-6R. In certain embodiments, the IL-6R inhibitor is tocilizumab. CRS can be identified based on clinical presentation. In some embodiments, other causes of fever, hypoxia, and hypotension are evaluated and treated.Laboratory tests can be used to monitor disseminated intravascular coagulation, hematological parameters, and pulmonary, cardiac, renal, and hepatic function. CRS can be managed according to the recommendations in Table 12. Procedures may include administering anti-seizure prophylaxis with levetiracetam to patients with CRS. In some embodiments, procedures include monitoring patients experiencing grade 2 or higher CRS (e.g., hypotension unresponsive to fluids or hypoxia requiring supplemental oxygenation) with continuous cardiac telemetry and pulse oximetry. In some embodiments, critical care level monitoring and supportive therapy may be used for severe or life-threatening CRS.For CRS refractory to first-line interventions such as tocilizumab or tocilizumab and corticosteroids, procedures include alternative treatment options (i.e., higher doses of corticosteroids, alternative anti-cytokine agents, e.g., anti-IL-1 and / or anti-TNF-α, anti-T-cell therapies). Refractory CRS is characterized by fever, end-organ toxicity (e.g., hypoxia, hypotension) that does not improve within 12 hours of first-line interventions, or the development of HLH / MAS.

[0157] In certain embodiments, the CAR-T cell treatment method further includes treating the subject with a pre-infusion drug comprising an antipyretic and an antihistamine for up to 1 hour prior to CAR-T cell administration. In certain embodiments, the antipyretic comprises either paracetamol or acetaminophen. In certain embodiments, the antipyretic is administered to the subject either orally or intravenously. In certain embodiments, the antipyretic is administered to the subject at a dose between 650 mg and 1000 mg. In certain embodiments, the antihistamine comprises diphenhydramine. In certain embodiments, the antihistamine is administered to the subject either orally or intravenously. In certain embodiments, the antihistamine is administered at a dose between 25 mg and 50 mg or its equivalent.The composition, comprising host cells expressing the CAR-coding nucleic acid sequences disclosed herein, or a vector comprising the CAR-coding nucleic acid sequences disclosed herein, can be administered to a mammal using standard administration techniques, including oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. The composition is preferably suitable for parenteral administration. The term "parenteral," as used herein, includes intravenous, intramuscular, subcutaneous, rectal, vaginal, and intraperitoneal administration. Most preferably, the composition is administered to a mammal using peripheral systemic administration by intravenous, intraperitoneal, or subcutaneous injection.The composition is particularly preferably administered via intravenous infusion.

[0158] The composition comprising host cells expressing the CAR-coding nucleic acid sequences disclosed herein, or a vector comprising the CAR-coding nucleic acid sequences disclosed herein, may be administered with one or more additional therapeutic agents, which may be given to the mammal simultaneously. "Cover administration" means the administration of one or more additional therapeutic agents and the composition comprising the host cells or vectors disclosed herein, which are sufficiently close in time such that the CARs disclosed herein can enhance the effect of one or more additional therapeutic agents, or vice versa.In this respect, the composition comprising the host cells or vectors disclosed herein may be administered first, and the one or more additional therapeutic agents may be administered second, or vice versa.

[0159] A CAR-expressing cell as described herein and the at least one additional therapeutic agent can be administered simultaneously, in the same or separate compositions, or sequentially. For sequential administration, the CAR-expressing cell described herein can be administered first, and the additional agent can be administered second, or the order of administration can be reversed.

[0160] In certain embodiments, CAR-T cell administration is preceded by a lymphodepleting regimen. In certain embodiments, the lymphodepleting regimen precedes CAR-T cell administration by approximately 2 to 7 days. In certain embodiments, the lymphodepleting regimen is administered intravenously. In certain embodiments, the lymphodepleting regimen includes the administration of cyclophosphamide or fludarabine. In certain embodiments, the cyclophosphamide is administered intravenously at a rate of 300 mg / m². 2 administered. In certain formulations, fludarabine is administered intravenously at a rate of 30 mg / m². 2 administered.

[0161] In certain embodiments, a lymphodepleting regimen is used that includes cyclophosphamide administered intravenously at 300 mg / m². 2 is administered, and fludarabine, which is given intravenously at 30 mg / m² 2The administration of CAR-T cells is preceded by approximately 2 days to approximately 7 days.

[0162] In certain embodiments, the subject further receives bridge therapy, wherein the bridge therapy comprises a short course of treatment with at least one bridge drug between apheresis and the lymphodepleting regimen, and wherein the at least one bridge drug had previously achieved a stable disease, minimal response, partial response, very good partial response, complete response, or stringent complete response for the subject. In certain embodiments, the subject exhibited an increase in tumor burden despite the bridge therapy. In certain embodiments, the subject exhibited an increase in tumor burden of approximately 25% or more despite the bridge therapy. Suitable bridge therapies include, for example, dexamethasone, daratumumab, bortezomib, cyclophosphamide, and pomalidomide.In some embodiments, the bridge therapy comprises pomalidomide, bortezomib, dexamethasone, daratumumab, or any combination thereof. In some embodiments, the bridge therapy comprises dexamethasone. In some embodiments, the bridge therapy comprises daratumumab. In some embodiments, the bridge therapy comprises bortezomib. In some embodiments, the bridge therapy comprises cyclophosphamide. In some embodiments, the bridge therapy comprises pomalidomide. In some embodiments, the bridge therapy comprises pomalidomide, bortezomib, and dexamethasone. In some embodiments, the bridge therapy comprises daratumumab, pomalidomide, and dexamethasone. In some embodiments, the subject received the bridge therapy from approximately every 10 days to approximately every 40 days. In some embodiments, the subject received the bridge therapy from approximately every 20 days to approximately every 30 days.In some embodiments, the subject received bridge therapy approximately every 21 days. In some embodiments, the subject received bridge therapy approximately every 15 days. In some embodiments, the subject received bridge therapy approximately every 25 days. In some embodiments, the subject received bridge therapy approximately every 21 days. In some embodiments, the subject received bridge therapy approximately every 28 days. In some embodiments, the subject received bridge therapy approximately every 30 days. In some embodiments, the subject received bridge therapy approximately every 35 days. In some embodiments, the subject received at least one, two, three, four, or more bridge therapies. In some embodiments, the subject received at least one bridge therapy. In some embodiments, the subject received at least two bridge therapies.In some embodiments, the subject has received at least three bridge therapies. In some embodiments, the subject has received at least four bridge therapies. In some embodiments, the subject has received at least five bridge therapies. In some embodiments, the subject has received at least six bridge therapies.

[0163] In certain embodiments, the subject is treated with pre-administration medication comprising an antipyretic and an antihistamine for up to approximately 1 hour prior to CAR-T cell delivery. In certain embodiments, the antipyretic comprises either acetaminophen or acetaminophen. In certain embodiments, the antipyretic is administered to the subject either orally or intravenously. In certain embodiments, the antipyretic is administered to the subject at a dose between 650 mg and 1000 mg. In certain embodiments, the antihistamine comprises diphenhydramine. In certain embodiments, the antihistamine is administered to the subject either orally or intravenously. In certain embodiments, the antihistamine is administered at a dose between 25 mg and 50 mg or its equivalent.In certain embodiments, the antipyretic comprises either paracetamol or acetaminophen and the antipyretic is administered to the subject either orally or intravenously in a dose between 650 mg and 1000 mg, and wherein the antihistamine comprises diphenhydramine and the antihistamine is administered to the subject either orally or intravenously in a dose between 25 mg and 50 mg or its equivalent.

[0164] In some embodiments, the procedures prior to CAR-T cell administration include the administration of a lymphodepleting chemotherapy regimen containing cyclophosphamide 300 mg / m² 2 intravenous (IV) and fludarabine 30-50 mg / m² 2 IV daily for 3 days, and the administration of pre-infusion drugs including an antipyretic (such as oral or intravenous acetaminophen 650 to 1000 mg) and antiphorales or 25-hydramine (such as intravenous diphenor-hydramine), wherein The CAR-T cells are administered 2-4 days after completion of lymphodepleting chemotherapy and The CAR-T cells are administered 30-60 minutes after the administration of the pre-infusion drugs.

[0165] In some embodiments, CAR-T cell administration is not given or is delayed if the patient has any of the following conditions: clinically significant active infection or inflammatory disorder; or non-hematological toxicities of grade ≥3 from cyclophosphamide and fludarabine conditioning, except for grade 3 nausea, vomiting, diarrhea, or constipation. CAR-T cell administration should be delayed until these events resolve to grade ≤1. In some embodiments, prophylactic systemic corticosteroids are not given.

[0166] In some embodiments, the method further includes diagnosing the subject for cytokine release syndrome (CRS). In preferred embodiments, the diagnosis is made according to the American Society of Transplantation and Cellular Therapy (ASTCT), formerly the American Society for Blood and Marrow Transplantation (ASBMT) Consensus Grading. A non-restrictive summary of the ASTCT consensus assessment for CRS diagnosis is given in Table 13.

[0167] In some embodiments, the method further includes treating the subject for cytokine release syndrome (CRS). In some embodiments, CRS is treated with an antipyretic. In some examples, CRS is treated with anticytokine therapy. In some embodiments, CRS is treated more than approximately 3 days after infusion. In some embodiments, CRS is treated without significantly reducing CAR-T cell expansion in vivo. In certain embodiments, the method further includes treating the subject for cytokine release syndrome more than approximately 3 days after CAR-T cell administration without significantly reducing CAR-T cell expansion in vivo. In some embodiments, CRS treatment includes administering an IL-6R inhibitor to the subject. In some embodiments, the IL-6R inhibitor is an antibody.In some embodiments, the antibody inhibits IL-6R by binding to its extracellular domain. In some embodiments, the IL-6R inhibitor prevents IL-6 from binding to IL-6R. In some embodiments, the IL-6R inhibitor is tocilizumab. In some embodiments, anti-cytokine therapy includes the administration of tocilizumab. In some embodiments, anti-cytokine therapy includes the administration of steroids. In some embodiments, the treatment of CRS includes treatment with monoclonal antibodies other than tocilizumab. In some embodiments, the antibodies other than tocilizumab target cytokines. In some embodiments, the cytokine targeted by the antibodies other than tocilizumab is IL-1. In some embodiments, the IL-1-targeting antibody is anakinra.In some embodiments, the cytokine targeted by antibodies other than tocilizumab is TNFα. In some embodiments, treatment for CRS includes administering a corticosteroid to the subject. In some embodiments, treatment for CRS includes using a vasopressor. In some embodiments, treatment for CRS includes intubation or mechanical ventilation. In some embodiments, treatment for CRS includes administering cyclophosphamide to the subject. In some embodiments, treatment for CRS includes administering etanercept to the subject. In some embodiments, treatment for CRS includes administering levetiracetam to the subject. In some embodiments, treatment for CRS includes supportive care.

[0168] In some embodiments, the method further includes diagnosing the subject for immune cell effector-associated neurotoxicity (ICANS). In some embodiments, the diagnosis is performed according to the National Cancer Institute Common Terminology Criteria for Negative Events (NCI CTCAE). In some embodiments, the diagnosis is performed according to the NCI CTCAE criteria, version 5.0. In some embodiments, the diagnosis is performed according to the American Society of Transplantation and Cellular Therapy (ASTCT) consensus evaluation system. In some embodiments, there is neurotoxicity consistent with ICAN. A non-restrictive summary of the ASTCT consensus evaluation system for ICANS diagnosis is given in Table 14. In some embodiments, the treatment of ICANS includes administering an IL-6R inhibitor to the subject.In some embodiments, the IL-6R inhibitor is an antibody. In some embodiments, the antibody inhibits IL-6R by binding to its extracellular domain. In some embodiments, the IL-6R inhibitor prevents IL-6 from binding to IL-6R. In some embodiments, the IL-6R inhibitor is tocilizumab. In some embodiments, the treatment of ICANS includes administering an IL-1 inhibitor to the subject. In some embodiments, the IL-1 inhibitor is an antibody. In a preferred aspect, the IL-1-inhibiting antibody is anakinra. In some embodiments, the treatment of ICANS includes administering a corticosteroid to the subject. In some embodiments, the treatment of ICANS includes administering levetiracetam to the subject. In some embodiments, the treatment of ICANS includes administering dexamethasone to the subject.In some embodiments, ICANS treatment includes administering methylprednisone sodium succinate to the subject. In some embodiments, ICANS treatment includes administering pethidine to the subject. In some embodiments, ICANS treatment includes administering one or more or all of tocilizumab, anakinra, a corticosteroid, levetiracetam, dexamethasone, methylprednisone sodium succinate, or pethidine.

[0169] If concurrent neurological toxicity is suspected during CRS or vice versa, procedures may include the administration of the following: • Corticosteroids according to the more aggressive intervention based on the CRS and neurological toxicity grades in Tables 1 and 2 of the approved label. • Tocilizumab according to CRS grade in Table 1 of the approved label. • Anti-seizure medication according to neurological toxicity in Table 2 of the approved label.

[0170] In some embodiments, the method further includes diagnosing the subject for cytopenias. In some embodiments, the cytopenias include one, several, or all of lymphopenia, neutropenia, and thrombocytopenia. Without being bound to any theory, grade 3 or 4 lymphopenia, but not grade 2 or lower, is defined by a lymphocyte count of less than 0.5 × 10⁻⁵. 9Grade 3 or 4 neutropenia, but not grade 2 or lower, is characterized by a neutrophil count of less than 1,000 cells per microliter of a subject's blood sample, and grade 3 or 4 thrombocytopenia, but not grade 2 or lower, is characterized by a platelet count of less than 50,000 cells per microliter of the subject's blood sample. In some embodiments, more than 75% of subjects with grade 3 or 4 lymphopenia recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 80% of subjects with grade 3 or 4 lymphopenia recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration.In some embodiments, more than 85% of subjects with grade 3 or 4 lymphopenia recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 90% of subjects with grade 3 or 4 lymphopenia recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 70% of subjects with grade 3 or 4 neutropenia recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 75% of subjects with grade 3 or 4 neutropenia recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration.In some embodiments, more than 80% of subjects with grade 3 or 4 neutropenia recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 85% of subjects with grade 3 or 4 neutropenia recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 30% of subjects with grade 3 or grade 4 thrombocytopenia recover to grade 2 or lower thrombocytopenia within 60 days of CAR-T cell administration. In some embodiments, more than 34% of subjects with grade 3 or grade 4 thrombocytopenia recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration.In some embodiments, more than 38% of subjects with grade 3 or grade 4 thrombocytopenia recover to grade 2 or lower 60 days after CAR-T cell administration. In some embodiments, more than 42% of subjects with grade 3 or grade 4 thrombocytopenia recover to grade 2 or lower 60 days after CAR-T cell administration.

[0171] Once the composition comprising host cells expressing the CAR-coding nucleic acid sequences disclosed herein, or a vector comprising the CAR-coding nucleic acid sequences disclosed herein, is administered to a mammal (e.g., a human), the biological activity of the CAR can be measured by any suitable prior art method. According to the methods disclosed herein, the CAR binds to BCMA on the multiple myeloma cells, and the multiple myeloma cells are destroyed. The binding of the CAR to BCMA on the surface of multiple myeloma cells can be tested using any suitable prior art method, including, for example, ELISA and flow cytometry.The ability of CAR to destroy multiple myeloma cells can be measured using any suitable, state-of-the-art method, such as cytotoxicity assays, as described, for example, in Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009) and Herman et al., J. Immunologische Methoden, 285(1): 25-40 (2004). The biological activity of CAR can also be measured by testing the expression of specific cytokines, such as CD107a, IFNγ, IL-2, and TNF.

[0172] The methods described herein can be used to treat various cancers, including both solid and liquid cancers. In certain embodiments, the methods are used to treat multiple myeloma. The methods described herein can be used as first-line, second-line, third-line, or combination therapy with other types of cancer therapies known in the art, such as chemotherapy, surgery, radiation therapy, gene therapy, immunotherapy, bone marrow transplantation, stem cell transplantation, targeted therapy, cryotherapy, ultrasound therapy, photodynamic therapy, radiofrequency ablation, or the like, in an adjuvant or neoadjuvant setting.

[0173] In certain embodiments, the cancer is multiple myeloma. In certain embodiments, the cancer is stage I, stage II, or stage III and / or stage A or stage B multiple myeloma, based on the Durie-Salmon staging system. In certain embodiments, the cancer is stage I, stage II, or stage III multiple myeloma based on the international staging system published by the International Myeloma Working Group (IMWG). In some embodiments, the multiple myeloma is progressive.

[0174] In certain aspects, the subject received prior treatment with one or more lines of therapy. In some embodiments, the number of prior therapy lines is 1. In certain embodiments, the number of prior therapy lines is 2. In some embodiments, the number of prior therapy lines is 3. In some embodiments, the number of prior therapy lines is 4. In some embodiments, the number of prior therapy lines is 5. In certain embodiments, prior therapy lines include surgery, radiotherapy, or autologous or allogeneic transplantation, or any combination of such treatments. In certain embodiments, the prior treatment includes treatment with a drug that is a proteasomal inhibitor (PI). Non-restrictive examples of a PI include bortezomib, carfilzomib, and ixazomib.In certain embodiments, the prior treatment includes treatment with a drug that is an immunomodulatory drug (IMiD). Non-restrictive examples of an IMiD include lenalidomide, pomalidomide, and thalidomide. In preferred embodiments, the prior treatment includes treatment with lenalidomide, optionally a treatment comprising lenalidomide and a proteasome inhibitor. In preferred embodiments, the subject received at least one prior line of therapy comprising lenalidomide, optionally lenalidomide and a proteasome inhibitor, and optionally one or two further prior lines of therapy. In certain embodiments, the prior treatment includes treatment with a drug that is a corticosteroid. Non-restrictive examples of a corticosteroid include dexamethasone and prednisone.In certain embodiments, the prior treatment includes treatment with a drug that is an alkylating agent. In certain embodiments, the prior treatment includes treatment with a drug that is an anthracycline. In certain embodiments, the prior treatment includes treatment with a drug that is an anti-CD38 antibody. Non-restrictive examples of an anti-CD38 antibody include daratumumab, isatuximab, and the investigational antibody TAK-079. In certain embodiments, the prior treatment includes treatment with a drug that is elotuzumab. In certain embodiments, the prior treatment includes treatment with a drug that is panobinostat.In certain embodiments, the prior treatment comprises treatment with at least one drug, wherein the at least one drug comprises at least one proteasome inhibitor (PI), one imivided de novo dilution (IMiD), and / or one anti-CD38 antibody. In certain embodiments, the prior line of therapy comprises treatment with at least one drug, wherein the at least one drug comprises at least one PI, one imivided de novo dilution (IMiD), and / or one alkylating agent. In certain embodiments, the subject relapses after the prior line of therapy.

[0175] In certain embodiments, the multiple myeloma is refractory to one or more or all of bortezomib, carfilzomib, ixazomib, lenalidomide, pomalidomide, thalidomide, dexamethasone, prednisone, alkylating agents, daratumumab, isatuximab, TAK-079, elotuzumab, and / or panobinostat. In certain embodiments, the multiple myeloma is refractory to at least one drug after one or more prior lines of therapy. In certain embodiments, the at least one drug to which the multiple myeloma is refractory includes an IMiD. In some embodiments, the IMiD includes lenalidomide, pomalidomide, or thalidomide. In some embodiments, the IMiD includes lenalidomide. In preferred embodiments, the multiple myeloma is lenalidomide-refractory multiple myeloma. In some embodiments, the IMiD is lenalidomide.In certain embodiments, multiple myeloma is refractory to at least two drugs after prior treatment. In certain embodiments, the at least two drugs for which the multiple myeloma is refractory include a proteasome inhibitor (PI) and an IMiD (e.g., lenalidomide). In certain embodiments, multiple myeloma is refractory to at least three drugs after the previous line of therapy. In certain embodiments, multiple myeloma is refractory to at least four drugs after the previous line of therapy. In certain embodiments, the at least four previous lines of therapy include treatment with at least one drug, wherein the at least one drug comprises at least one of a PI, an IMiD, an anti-CD38 antibody, and / or an alkylating agent.In certain cases, multiple myeloma is refractory to at least five drugs after the previous line of therapy.

[0176] In some embodiments, the subject has between approximately 10% and approximately 30% bone marrow plasma cells prior to the administration of the CAR-T cells.

[0177] In certain embodiments, bone marrow aspirate or biopsy can be performed for clinical assessments, or bone marrow aspirate can be performed for biomarker evaluations. In certain embodiments, clinical staging (morphology, cytogenetics and immunohistochemistry or immunofluorescence or flow cytometry) can be performed. In certain embodiments, a portion of the bone marrow aspirate can be immunophenotyped and monitored for BCMA, checkpoint ligand expression in CD138-positive multiple myeloma cells, and checkpoint expression on T cells. In certain embodiments, minimal residual disease (MRD) in subjects can be monitored using next-generation sequencing (NGS) of bone marrow aspirate DNA. NGS of bone marrow aspirate DNA is known to those skilled in the art. In certain embodiments, NGS is performed using clonoSeq.In certain embodiments, baseline bone marrow aspirates can be used to define the myeloma clones, and post-treatment samples can be used to assess MRD negativity. In certain embodiments, the MRD negativity status can be based on evaluable samples. In certain embodiments, evaluable samples are those that have passed one or more or all of the calibration, quality control, and cell sufficiency tests required for evaluation at a specific sensitivity level. In some embodiments, the sensitivity level is 10. -6 In certain embodiments, the sensitivity level is 10 -6 , the sensitivity level 10 -5 In certain embodiments, the sensitivity level is 10 -4 In certain embodiments, the sensitivity level is 10 -3 .

[0178] In certain embodiments, a subject's response to the treatment procedure is evaluated using the response criteria based on the International Myeloma Working Group (IMWG), which are summarized in Table 6. In certain embodiments, the response may be classified as a stringent complete response (sCR). In certain embodiments, the response may be classified as a complete response (CR) that is inferior to a stringent complete response (sCR). In certain embodiments, the response may be classified as a very good partial response (VGPR) that is inferior to a complete response (CR). In certain embodiments, the response may be classified as a partial response (PR) that is inferior to a very good partial response (VGPR).In certain embodiments, the reaction can be classified as a minimal response (MR), which is worse than a partial response (PR). In certain embodiments, the reaction can be classified as a stable disease (SD), which is worse than a minimal response (MR). In certain embodiments, the reaction can be classified as a progressive disease (PD), which is worse than a stable disease.

[0179] In certain embodiments, the tests used to assess the response criteria based on the International Myeloma Working Group (IMWG) include myeloma protein (M-protein) measurements in serum and urine, serum calcium corrected for albumin, bone marrow examination, skeletal overview, and documentation of extramedullary plasmacytomas.

[0180] Non-restrictive examples of tests for measuring M-protein in blood and urine are known to the average professional and include quantitative serum Ig, serum protein electrophoresis (SPEP), serum immunofixation electrophoresis, serum FLC assay, 24-hour urine M-protein quantification by electrophoresis (UPEP), urine immunofixation electrophoresis, and serum β2-microglobulin.

[0181] The calculation of serum calcium, corrected for albumin, in blood samples to detect hypercalcemia is well known to those skilled in the art. Without adhering to any specific theory, calcium binds to albumin, and only the unbound (free) calcium is biologically active; therefore, the serum calcium level must be adjusted to reflect abnormal albumin levels ("corrected serum calcium").

[0182] In certain embodiments, skeletal measurements of one or all of the skull, entire spine, pelvis, thorax, humeri, femora, and all other bones can be performed and evaluated either by radiographs (“X-rays”) or by low-dose diagnostic quality examinations using computed tomography (CT) without the use of IV contrast, both of which are known to the person skilled in the art. In certain embodiments, after T-cell administration and before disease progression is confirmed, radiographs or CT scans can be performed locally whenever clinically indicated based on symptoms to document the response or progression. In certain embodiments, magnetic resonance imaging (MRI) can be used to evaluate bone disease but does not replace skeletal measurements. MRI is known to the person skilled in the art.In certain embodiments, when a radionuclide bone scan is used in screening, both methods, in addition to full skeletal measurement, can be used to document the disease status. Radionuclide bone scans are known to those skilled in the art. In certain embodiments, the radionuclide bone scan and the full skeletal measurement can be performed simultaneously. In certain embodiments, a radionuclide bone scan may not replace a full skeletal measurement. In certain embodiments, if a subject exhibits disease progression manifested by symptoms of pain due to bone changes, the disease progression can be documented by skeletal measurement or other radiographs, depending on the symptoms experienced by the subject.

[0183] In certain embodiments, extramedullary plasmacytomas can be documented by clinical examination or MRI. In certain embodiments, extramedullary plasmacytomas can be documented by CT scan if there was no contraindication to the use of IV contrast. In certain embodiments, extramedullary plasmacytomas can be documented by a fusion of positron emission tomography (PET) and CT scans if the CT component has sufficient diagnostic quality. In certain embodiments, the assessment of measurable sites of extramedullary disease can be performed, measured, or evaluated locally for individuals every 4 weeks until the development of a confirmed complete response (CR) or confirmed disease progression. In certain embodiments, the evaluation of extramedullary plasmacytomas can be performed every 12 weeks.

[0184] In certain embodiments, to qualify for VGPR, PR, or MR, the sum of the products of the vertical diameters of the existing extramedullary plasmacytomas may be reduced by more than 90% or by at least 50%, respectively. In certain embodiments, to qualify for disease progression, either the sum of the products of the vertical diameters of the existing extramedullary plasmacytomas must be increased by at least 50%, or the longest diameter of the previous lesion must be > 1 cm in the short axis and increased by at least 50%, or a new plasmacytoma must have developed. In certain embodiments, the sum of the products of the vertical diameters of the reported plasmacytomas had to be increased by at least 50% to qualify for disease progression if not all existing extramedullary plasmacytomas were reported.In certain embodiments, CR can be defined as the disappearance of the original M protein associated with multiple myeloma during immunofixation when the study treatment interferes with the immunofixation assay.

[0185] In certain embodiments, a subject's response to the treatment procedure is evaluated with respect to changes in disease burden or tumor burden. Disease burden or tumor burden represents the nature of the subject's measurable disease. In some embodiments, the change in tumor burden can be evaluated with respect to changes in paraprotein levels during treatment. In some embodiments, the paraprotein is an M-protein in serum. In some embodiments, the change in tumor burden is evaluated with respect to the difference between involved and uninvolved free light chain (dFLC). In some embodiments, the change in tumor burden is evaluated with respect to the maximum paraprotein reduction from baseline, i.e., before CAR-T cell administration.In some embodiments, the change in tumor burden is evaluated at a median follow-up of 28 days or more after CAR-T cell administration. In some embodiments, the change in tumor burden is evaluated at a median follow-up of 1 month or more after CAR-T cell administration. In some embodiments, the change in tumor burden is evaluated at a median follow-up of 3 months or more after CAR-T cell administration. In some embodiments, the change in tumor burden is evaluated at a median follow-up of 6 months or more after CAR-T cell administration. In some embodiments, the change in tumor burden is evaluated at a median follow-up of 9 months or more after CAR-T cell administration.In some embodiments, the change in tumor burden is evaluated at a median follow-up time of more than or equal to 12 months after administration of CAR-T cells.

[0186] In certain embodiments, the subject is retreated by administering a second intravenous infusion of a second dose of CAR-T cells. In certain embodiments, the retreatment dose comprises 1.0 × 10 5 up to 5.0 × 10 6 CAR-T cells per kilogram of the subject's mass. In certain embodiments, the post-treatment dose comprises approximately 0.75 × 10 5CAR-T cells per kilogram of the subject's mass. In certain embodiments, the subject is retreated after the onset of disease progression, following a best response of minimal response or better after the first CAR-T cell infusion. In certain embodiments, the time between the first CAR-T cell infusion and the detection of disease progression is at least six months.

[0187] In one aspect, a procedure is provided for the treatment of a subject who has multiple myeloma, wherein the procedure comprises administering a composition comprising a therapeutically effective number of T cells comprising a chimeric antigen receptor (CAR) to the subject via a single intravenous infusion to deliver a dose of CAR-expressing T cells (CAR-T cells) to the subject.

[0188] In some embodiments, the subject received prior treatment with at least one to three prior lines of therapy. In some embodiments, the prior line of therapy includes treatment with at least one drug, wherein the at least one drug comprises at least one proteasome inhibitor (PI), one imiD, and one anti-CD38 antibody. In some embodiments, the subject relapsed after the prior line of therapy.

[0189] In some embodiments, the multiple myeloma is refractory to at least two drugs after the previous line of therapy. In some embodiments, the at least two drugs to which the subject is refractory include a proteasome inhibitor (PI) and an IMiD (e.g., lenalidomide). In some embodiments, the multiple myeloma is refractory to at least three drugs after the previous line of therapy. In some embodiments, the multiple myeloma is refractory to at least four drugs after the previous line of therapy. In some embodiments, the multiple myeloma is refractory to at least five drugs after the previous line of therapy.

[0190] In some embodiments, the subject is older than 65 years. In some embodiments, the subject is Black or African American. In some embodiments, the subject has received 1-3 prior lines of therapy. In some embodiments, the subject has received at least 1 prior line of therapy. In some embodiments, the subject has received at least 2 prior lines of therapy. In some embodiments, the subject has received at least 3 prior lines of therapy. In some embodiments, the subject has received at least 4 prior lines of therapy. In some embodiments, the multiple myeloma or the subject is refractory to three classes of drugs, i.e., the multiple myeloma or the subject is triple-class refractory. In some embodiments, the multiple myeloma or the subject is refractory to five drugs or active substances.The multiple myeloma or the subject is pentadrug-refractory. In some embodiments, the subject has high-risk factors, including high-risk cytogenetic abnormalities, soft tissue plasmacytomas, refractory triple-class or other high-risk factors. In some embodiments, the subject has standard-risk cytogenetics. In some embodiments, the subject has high-risk cytogenetics. In some embodiments, the cytogenetic abnormality is a standard-risk cytogenetic abnormality. In some embodiments, the cytogenetic abnormality is a high-risk cytogenetic abnormality. In some embodiments, the subject has one or more high-risk cytogenetic abnormalities selected from a group that includes Gain / amp(1q), del(17p), t(4;14), t(14;16), or any combination thereof.In some embodiments, the cytogenetic anomaly comprises Gain / amp(1q). In some embodiments, the cytogenetic anomaly comprises del(17p). In some embodiments, the cytogenetic anomaly comprises t(4;14). In some embodiments, the cytogenetic anomaly comprises t(14;16). In some embodiments, the subject has two, three, four, five, or more cytogenetic anomalies. In some embodiments, the subject has at least two cytogenetic anomalies. In some embodiments, the subject has at least three cytogenetic anomalies. In some embodiments, the subject has at least four cytogenetic anomalies. In some embodiments, the subject has at least five cytogenetic anomalies. In some embodiments, the subject has at least six cytogenetic anomalies. In some embodiments, the subject has at least seven cytogenetic anomalies.In some embodiments, the subject or multiple myeloma was characterized as stage III according to the International Staging System. In some embodiments, the subject has soft tissue plasmacytomas. In some embodiments, the subject has bone marrow plasma cells between approximately 10% and approximately 30% prior to CAR-T cell administration. In some embodiments, the subject has bone marrow plasma cells between approximately 31% and approximately 59% prior to CAR-T cell administration. In some embodiments, the subject has bone marrow plasma cells between approximately 60% and approximately 100% prior to CAR-T cell administration. In some embodiments, the subject has BCMA expression in the tumor that is lower than the median in a population of patients with multiple myeloma or in any randomly selected population.In some embodiments, the subject exhibits BCMA expression in the tumor that is greater than or equal to the median in a population of patients with multiple myeloma or in any randomly selected population. In some embodiments, plasmacytomas are present in the subject. In some embodiments, the plasmacytomas are bone-based. In some embodiments, the plasmacytomas are extramedullary. In some embodiments, the plasmacytomas are both bone-based and extramedullary.

[0191] In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject between approximately 1% and approximately 100%, between approximately 60% and approximately 100%, between approximately 65% ​​and approximately 100%, between approximately 70% and approximately 100%, between approximately 75% and approximately 100%, between approximately 80% and approximately 100%, between approximately 85% and approximately 100%, between approximately 90% and approximately 100%, between approximately 92% and approximately 100%, between approximately 95% and approximately 100%, between approximately 96% and approximately 100%, between approximately 97% and approximately 100%, between approximately 98% and approximately 100%, or between approximately 99% and approximately 100%. In certain embodiments, the treatment procedure is effective in maintaining a reduced tumor burden in the subject of approximately 100%.In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject of between approximately 1% and approximately 100% at a rate of between approximately 1% and approximately 100%. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject of between approximately 60% and approximately 100% at a rate of between approximately 1% and approximately 100%. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject of between approximately 65% ​​and approximately 100% at a rate of between approximately 1% and approximately 92%. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject of between approximately 70% and approximately 100% at a rate of between approximately 1% and approximately 88%.In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject of between approximately 90% and approximately 100% at a rate of between approximately 1% and approximately 88%. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject of between approximately 95% and approximately 100% at a rate of between approximately 1% and approximately 88%. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject of between approximately 99% and approximately 100% at a rate of between approximately 1% and approximately 88%. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject of approximately 100% at a rate of between approximately 1% and approximately 83%.

[0192] In certain embodiments, the treatment method is effective in maintaining a negative minimal residual disease (MRD) status in the subject. In certain embodiments, the treatment method is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -6 in the subject. In certain embodiments, the treatment procedure is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -5 in the subject. In certain embodiments, the treatment procedure is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -4in the subject. In certain embodiments, the treatment procedure is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -3in the subject. In certain embodiments, the treatment procedure is effective in maintaining an MRD-negative status when evaluated in bone marrow. In certain embodiments, the treatment procedure is effective in maintaining an MRD-negative status when evaluated using a bone marrow sample that is evaluable. In certain embodiments, the treatment procedure is effective in maintaining an MRD-negative status when evaluated using bone marrow DNA.In some embodiments, the method is effective in maintaining a negative minimal residual disease (MRD) status in the subject, as assessed in the bone marrow at a follow-up period of approximately 28 days or later after CAR-T cell administration, approximately 2 months or later after CAR-T cell administration, approximately 3 months or later after CAR-T cell administration, approximately 6 months or later after CAR-T cell administration, approximately 9 months or later after CAR-T cell administration, or approximately 12 months or later after CAR-T cell administration. In some embodiments, the negative minimal residual disease (MRD) status is maintained at an initial follow-up period between approximately 28 days and approximately 179 days after CAR-T cell infusion.

[0193] In certain embodiments, the treatment method is effective in maintaining an initial minimal residual disease (MRD) negative status in the subject. In certain embodiments, the treatment method is effective in maintaining the MRD-negative status at a sensitivity level of 10. -5 In certain embodiments, the treatment method is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -6 in the subject. In certain embodiments, the treatment procedure is effective in maintaining the MRD-negative status at a sensitivity level of 10. -4 In certain embodiments, the treatment method is effective in maintaining the MRD-negative status at a sensitivity level of 10 -3In certain embodiments, the treatment procedure is effective in maintaining MRD-negative status when evaluated using a bone marrow sample. In certain embodiments, the treatment procedure is effective in maintaining MRD-negative status when evaluated using an evaluable bone marrow sample. In certain embodiments, the treatment procedure is effective in maintaining MRD-negative status when evaluated using bone marrow DNA.In some embodiments, the method is effective in maintaining a negative minimal residual disease (MRD) status in the subject as assessed in bone marrow at a second follow-up period between approximately 29 and 359 days after CAR-T cell administration, between approximately 29 and 9 months after CAR-T cell administration, between approximately 29 and 6 months after CAR-T cell administration, between approximately 29 and 3 months after CAR-T cell administration, or between approximately 29 and 2 months after CAR-T cell administration. In some embodiments, the method is effective in maintaining a negative minimal residual disease (MRD) status in the subject as assessed in bone marrow at a second follow-up period between approximately 180 and 359 days after CAR-T cell infusion.In some embodiments, the method is effective in maintaining the negative minimal residual disease (MRD) status in the subject, as assessed in the bone marrow at a second follow-up time between approximately 360 days and approximately 539 days after CAR-T cell infusion.

[0194] In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status. In certain embodiments, the efficacy of the treatment procedure is assessed by assessing the proportion of subjects with MRD-negative status at a sensitivity level of 10. -6 In certain embodiments, the effectiveness of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status at a sensitivity level of 10. -5In certain embodiments, the effectiveness of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status at a sensitivity level of 10. -4 In certain embodiments, the effectiveness of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status at a sensitivity level of 10. -3In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of individuals with MRD-negative status at a median follow-up time between CAR-T cell administration and approximately 359 days post-administration, between CAR-T cell administration and approximately 9 months post-administration, between CAR-T cell administration and approximately 6 months post-administration, between CAR-T cell administration and approximately 3 months post-administration, between CAR-T cell administration and approximately 2 months post-administration, or between CAR-T cell administration and approximately 29 days post-administration.In some embodiments, the method is effective in maintaining the negative status of minimal residual disease (MRD) at a rate of approximately 44% or less at a sensitivity threshold of 10. -5 With a follow-up period of approximately 12 months after the CAR-T cell infusion, a rate of approximately 55% was observed at a sensitivity threshold of 10. -5 With a follow-up period of approximately 12 months after administration of the CAR-T cells, a rate of approximately 65% ​​or less at a sensitivity threshold of 10 -5 With a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 57% or less at a sensitivity threshold of 10 -4 With a follow-up period of approximately 18 months after the CAR-T cell infusion, a rate of approximately 67% was observed at a sensitivity threshold of 10. -4With a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 76% or less at a sensitivity threshold of 10 -4 With a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 47% or less at a sensitivity threshold of 10 -5 With a follow-up period of approximately 18 months after the CAR-T cell infusion, a rate of approximately 58% was observed at a sensitivity threshold of 10. -5 With a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 68% or less at a sensitivity threshold of 10 -5 With a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 29% or less at a sensitivity threshold of 10 -6With a follow-up period of approximately 18 months after the CAR-T cell infusion, a rate of about 39% was observed at a sensitivity threshold of 10. -6 with a follow-up period of approximately 18 months after the CAR-T cell infusion, or a rate of approximately 50% or less at a sensitivity threshold of 10 -6 with a follow-up period of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in maintaining a negative minimal residual disease (MRD) status at a rate between approximately 44% and approximately 65% ​​at a sensitivity threshold of 10. -5 With a follow-up period of approximately 12 months after the CAR-T cell infusion, a rate between approximately 57% and approximately 76% was observed at a sensitivity threshold of 10. -4With a follow-up period of approximately 18 months after the CAR-T cell infusion, a rate between approximately 47% and approximately 68% was observed at a sensitivity threshold of 10. -5 with a follow-up period of approximately 18 months after the CAR-T cell infusion, or a rate between approximately 29% and approximately 50% at a sensitivity threshold of 10 -6 with a follow-up period of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in maintaining a negative minimal residual disease (MRD) status at a rate of approximately 55% at a sensitivity threshold of 10. -5 With a follow-up period of approximately 12 months after administration of the CAR-T cells, a rate of approximately 67% was observed at a sensitivity threshold of 10. -4With a follow-up period of approximately 18 months after the CAR-T cell infusion, a rate of about 58% was observed at a sensitivity threshold of 10. -5 with a follow-up period of approximately 18 months after the CAR-T cell infusion, or a rate of approximately 39% at a sensitivity threshold of 10 -6 with a follow-up period of approximately 18 months after the infusion of CAR-T cells.

[0195] In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of subjects with evaluable bone marrow and MRD-negative status. In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of subjects with evaluable bone marrow and MRD-negative status at a sensitivity level of 10. -6In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a sensitivity level of 10. -5 In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a sensitivity level of 10. -4 In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a sensitivity level of 10. - 3In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a median follow-up time between CAR-T cell administration and approximately 359 days post-administration, between CAR-T cell administration and approximately 9 months post-administration, between CAR-T cell administration and approximately 6 months post-administration, between CAR-T cell administration and approximately 3 months post-administration, between CAR-T cell administration and approximately 2 months post-administration, or between CAR-T cell administration and approximately 29 days post-administration.In some embodiments, the method is effective in maintaining the negative status of minimal residual disease (MRD) at a rate of approximately 83% or less in subjects with evaluable samples at a sensitivity threshold of 10. -5 With a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 93% was observed in subjects with evaluable samples at a sensitivity threshold of 10. -5 with a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 98% or less in subjects with evaluable samples at a sensitivity threshold of 10 -5 With a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 82% or less in subjects with evaluable samples at a sensitivity threshold of 10 -5With a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 92% in subjects with evaluable samples at a sensitivity threshold of 10 -5 with a follow-up period of approximately 18 months after CAR-T cell infusion, or a rate of approximately 97% or less in subjects with evaluable samples at a sensitivity threshold of 10 -5 with a follow-up period of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in maintaining a negative minimal residual disease (MRD) status at a rate between approximately 83% and approximately 98% in subjects with evaluable samples at a sensitivity threshold of 10. -5with a follow-up period of approximately 12 months after CAR-T cell infusion, or with a rate between approximately 82% and approximately 97% in subjects with evaluable samples at a sensitivity threshold of 10 -5 with a follow-up period of approximately 12 months after CAR-T cell infusion, or with a rate between approximately 82% and approximately 97% in subjects with evaluable samples at a sensitivity threshold of 10 -5 with a follow-up period of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in maintaining a negative minimal residual disease (MRD) status at a rate of approximately 93% in subjects with evaluable samples at a sensitivity threshold of 10. -5with a follow-up period of approximately 12 months after CAR-T cell infusion, or with a rate of approximately 92% in subjects with evaluable samples at a sensitivity threshold of 10 -5 with a follow-up period of approximately 18 months after the infusion of CAR-T cells.

[0196] In some embodiments, the method is effective in obtaining at least one response in the subject after infusion of the CAR-T cells, wherein the at least one response comprises, in order from best to worst, a stringent complete response, a complete response, a very good partial response, a partial response, or a minimal response.

[0197] In some embodiments, the method is effective in obtaining a first response within approximately 27 days or later, approximately 29 days or later, approximately 42 days or later, approximately 89 days or later, or approximately 321 days or later after CAR-T cell infusion. In some embodiments, the method is effective in obtaining a first response between approximately 27 and approximately 321 days after CAR-T cell infusion. In some embodiments, the method is effective in obtaining a first response between approximately 27 and approximately 89 days after CAR-T cell infusion. In some embodiments, the method is effective in obtaining a first response approximately 42 days after CAR-T cell infusion. In some embodiments, the method is effective in obtaining a first response approximately 29 days after CAR-T cell infusion.

[0198] In certain embodiments, the effectiveness of the treatment method is evaluated by assessing the proportion of subjects with a stringent-complete response. In certain embodiments, the effectiveness of the treatment method is assessed by assessing the proportion of subjects with a complete response or better. In certain embodiments, the effectiveness of the treatment method is assessed by assessing the proportion of subjects with a very good partial response or better. In certain embodiments, the effectiveness of the treatment method is assessed by assessing the proportion of subjects with a partial response or better. In certain embodiments, the effectiveness of the treatment method is assessed by assessing the proportion of individuals with a minimal response or better.

[0199] In some embodiments, the method is effective in obtaining a best response of minimal response, partial response, very good partial response, complete response, or stringently complete response—that is, a best response of minimal response or better. In some embodiments, the rate at which the method is effective in obtaining a best response of minimal response or better is referred to as the clinical benefit rate.In some embodiments, the method is effective in obtaining the best response from a minimum response or better with a rate of approximately 91% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 97% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate between approximately 99% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 93% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 98% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, or a rate between approximately 100% or less at a follow-up period of approximately 18 months after CAR-T cell infusion.In some embodiments, the method is effective in obtaining the best response from a minimal response, partial response, very good partial response, complete response, or stringent complete response at a rate between approximately 91% and approximately 99% at a follow-up time of approximately 12 months after CAR-T cell infusion, or at a rate between approximately 93% and approximately 100% at a follow-up time of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response from a minimal response, partial response, very good partial response, complete response, or stringent complete response at a rate of approximately 97% at a follow-up time of approximately 12 months after CAR-T cell infusion, or at a rate of approximately 98% at a follow-up time of approximately 18 months after CAR-T cell infusion.

[0200] In some embodiments, the method is effective in obtaining a stringently complete reaction at a rate of about 40% to about 90%. In some embodiments, the method is effective in obtaining a stringently complete reaction at a rate of about 50% to about 80%. In some embodiments, the method is effective in obtaining a stringently complete reaction at a rate of about 58.2%. In some embodiments, the method is effective in obtaining a stringently complete reaction at a rate of about 68.8%.

[0201] In some embodiments, the method is effective in obtaining a complete reaction at a rate of about 10% to about 20%. In some embodiments, the method is effective in obtaining a complete reaction at a rate of about 14.9%. In some embodiments, the method is effective in obtaining a complete reaction at a rate of about 17.6%.

[0202] In some embodiments, the method is effective in obtaining a very good partial response.

[0203] In some embodiments, the method is effective in obtaining a partial response.

[0204] In some embodiments, the method is effective in obtaining a best-in-class partial reaction, a very good partial reaction, a complete reaction, or a stringently complete reaction, i.e., a best-in-class partial reaction or better. In some embodiments, the rate at which the method is effective in obtaining a best-in-class partial reaction or better is called the overall survival rate or overall reaction rate.In some embodiments, the method is effective in obtaining a best partial response or better with a rate of approximately 91% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 97% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 99% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 93% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 97% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, or a rate of approximately 100% or less at a follow-up period of approximately 18 months after CAR-T cell infusion.In some embodiments, the method is effective in obtaining the best response from a partial response, very good partial response, complete response, or stringent complete response at a rate between approximately 91% and approximately 99% at a follow-up period of approximately 12 months after CAR-T cell infusion, or at a rate between approximately 93% and approximately 100% at a follow-up period of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response from a partial response, very good partial response, complete response, or stringent complete response at a rate of approximately 97% at a follow-up period of approximately 12 months after CAR-T cell infusion, or at a rate of approximately 97% at a follow-up period of approximately 18 months after CAR-T cell infusion.

[0205] In some embodiments, the method is effective in obtaining a best response of very good partial response, complete response or stringent complete response, i.e. a best response of very good partial response or better.In some embodiments, the method is effective in obtaining the best response of very good partial response or better, at a rate of about 86% or less at a follow-up time of about 12 months after CAR-T cell infusion, a rate of about 93% or less at a follow-up time of about 12 months after CAR-T cell infusion, a rate of about 97% or less at a follow-up time of about 12 months after CAR-T cell infusion, a rate of about 88% or less at a follow-up time of about 18 months after CAR-T cell infusion, a rate of about 95% or less at a follow-up time of about 18 months after CAR-T cell infusion, or a rate of about 98% or less at a follow-up time of about 18 months after CAR-T cell infusion.In some embodiments, the method is effective in obtaining the best response from a very good partial response, a complete response, or a stringent complete response at a rate between approximately 86% and approximately 97% at a follow-up time of approximately 12 months after CAR-T cell infusion, or at a rate between approximately 88% and approximately 98% at a follow-up time of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response from a very good partial response, a complete response, or a stringent complete response at a rate of approximately 93% at a follow-up time of approximately 12 months after CAR-T cell infusion, or at a rate of approximately 95% at a follow-up time of approximately 18 months after CAR-T cell infusion.

[0206] In some embodiments, the method is effective in obtaining a best-of-complete response or stringent-complete response, i.e., a best-of-complete response or better.In some embodiments, the method is effective in obtaining the best response of complete response or better at a rate of approximately 57% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 67% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 76% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 73% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 83% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, or a rate of approximately 89% or less at a follow-up period of approximately 18 months after CAR-T cell infusion.In some embodiments, the method is effective in obtaining the best response of complete response or stringent complete response at a rate between approximately 57% and approximately 76% at a follow-up time of approximately 12 months after CAR-T cell infusion, or at a rate between approximately 73% and approximately 89% at a follow-up time of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response of complete response or stringent complete response at a rate of approximately 67% at a follow-up time of approximately 12 months after CAR-T cell infusion, or at a rate of approximately 83% at a follow-up time of approximately 18 months after CAR-T cell infusion.

[0207] In some embodiments, the method is effective in obtaining a best response of stringent-complete response.In some embodiments, the method is effective in obtaining the best response of stringent complete response with a rate of approximately 57% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 67% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 76% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 73% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 83% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, or a rate of approximately 89% or less at a follow-up period of approximately 18 months after CAR-T cell infusion.In some embodiments, the method is effective in obtaining the best response of stringent complete response at a rate between approximately 57% and approximately 76% at a follow-up time of approximately 12 months after CAR-T cell infusion, or at a rate between approximately 73% and approximately 89% at a follow-up time of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response of stringent complete response at a rate of approximately 67% at a follow-up time of approximately 12 months after CAR-T cell infusion, or at a rate of approximately 83% at a follow-up time of approximately 18 months after CAR-T cell infusion.

[0208] In some embodiments, the method is effective in obtaining the best response within approximately 27 days or later, 78 days or later, 153 days or later, 293 days or later, or approximately 534 days or later after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response before a time between approximately 27 days and approximately 534 days after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response before a time between approximately 27 days and approximately 293 days after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response before approximately 153 days after CAR-T cell infusion. In some embodiments, the method is effective in obtaining the best response before approximately 78 days after CAR-T cell infusion.

[0209] In some embodiments, the method is effective in maintaining a response in the subject at follow-up times between the time of the first response and approximately 180 days after CAR-T cell infusion, between the time of the first response and approximately 357 days after CAR-T cell infusion, between the time of the first response and approximately 606 days after CAR-T cell infusion, or between the time of the first response and approximately 654 days after CAR-T cell infusion. In some embodiments, the method is effective in maintaining a response at a rate of approximately 77% or less at a follow-up time of approximately 6 months after CAR-T cell infusion, and at a rate of approximately 85% or less at a follow-up time of approximately 6 months after CAR-T cell infusion.a rate of approximately 91% or less at a follow-up period of approximately 6 months after CAR-T cell infusion, a rate of approximately 63% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 74% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 81% or less at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 56% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 67% or less at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 75% or less at a follow-up period of approximately 18 months after CAR-T cell infusion,a rate of approximately 52% or less at a follow-up period of approximately 21 months after CAR-T cell infusion, a rate of approximately 63% or less at a follow-up period of approximately 21 months after CAR-T cell infusion, a rate of approximately 72% or less at a follow-up period of approximately 21 months after CAR-T cell infusion, a rate of approximately 48% or less at a follow-up period of approximately 24 months after CAR-T cell infusion,a response rate of approximately 60% or less at a follow-up period of approximately 24 months after CAR-T cell infusion, or a response rate of approximately 70% or less at a follow-up period of approximately 24 months after CAR-T cell infusion. In some embodiments, the method is effective in maintaining a response at a rate between approximately 77% and approximately 91% at a follow-up period of approximately 6 months after CAR-T cell infusion, a rate between approximately 63% and approximately 81% at a follow-up period of approximately 12 months after CAR-T cell infusion, and a rate between approximately 56% and approximately 75% at a follow-up period of approximately 18 months after CAR-T cell infusion.a rate between approximately 52% and approximately 72% at a follow-up period of approximately 21 months after CAR-T cell infusion, or a rate between approximately 48% and approximately 70% at a follow-up period of approximately 24 months after CAR-T cell infusion. In some embodiments, the method is effective in maintaining a response at a rate of approximately 85% at a follow-up period of approximately 6 months after CAR-T cell infusion, a rate of approximately 74% at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 67% at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 63% at a follow-up period of approximately 21 months after CAR-T cell infusion, or a rate of approximately 60% at a follow-up period of approximately 24 months after CAR-T cell infusion.

[0210] In some embodiments, wherein the method is effective in maintaining a negative minimal residual disease (MRD) status in the subject, which is expressed in the bone marrow at a sensitivity threshold of 10 -5The response is evaluated between the time of CAR-T cell administration and approximately 3 months after CAR-T cell administration. In some embodiments, the method is effective in obtaining either a negative complete minimal residual disease (MRD) response or a negative stringent complete MRD response at a rate of approximately 25% or less at a follow-up of approximately 12 months after CAR-T cell infusion, a rate of approximately 34% or less at a follow-up of approximately 12 months after CAR-T cell infusion, a rate of approximately 44% or less at a follow-up of approximately 12 months after CAR-T cell infusion, a rate of approximately 33% or less at a follow-up of approximately 18 months after CAR-T cell infusion, and a rate of approximately 43% or less at a follow-up of approximately 18 months.months after CAR-T cell infusion, or a rate of approximately 54% or less at a follow-up period of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in obtaining either a negative complete minimal residual disease (MRD) response or a negative stringent complete MRD response at a rate between approximately 25% and approximately 44% at a follow-up period of approximately 12 months after CAR-T cell infusion, or a rate between approximately 33% and approximately 54% at a follow-up period of approximately 18 months after CAR-T cell infusion. In some embodiments, the method is effective in obtaining either a negative complete MRD response or a negative stringent complete MRD response at a rate of approximately 34% at aFollow-up period of approximately 12 months after CAR-T cell infusion or with a rate of approximately 43% with a follow-up period of approximately 18 months after CAR-T cell infusion.

[0211] In some embodiments, the method is effective in maintaining progression-free survival in the subject. In some embodiments, the method is effective in maintaining progression-free survival in the subject at a time between CAR-T cell infusion and approximately 209 days post-infusion, between CAR-T cell infusion and approximately 386 days post-infusion, between CAR-T cell infusion and approximately 632 days post-infusion, or between CAR-T cell infusion and approximately 684 days post-infusion. In some embodiments, the method is effective in maintaining progression-free survival at a rate of approximately 79% or higher at a follow-up period of approximately 6 months post-infusion, or at a rate of approximately 88% or higher at a follow-up period of approximately 6 months post-infusion.a rate of approximately 93% or more at a follow-up period of approximately 6 months after CAR-T cell infusion, a rate of approximately 67% or more at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 76% or more at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 84% or more at a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 57% or more at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 67% or more at a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 75% or more at a follow-up period of approximately 18 months after CAR-T cell infusion,a rate of approximately 57% or more at a follow-up period of approximately 21 months after CAR-T cell infusion, a rate of approximately 67% or more at a follow-up period of approximately 21 months after CAR-T cell infusion, a rate of approximately 75% or more at a follow-up period of approximately 21 months after CAR-T cell infusion, a rate of approximately 49% or more at a follow-up period of approximately 24 months after CAR-T cell infusion,a rate of approximately 61% or more with a follow-up period of approximately 24 months after CAR-T cell infusion, or a rate of approximately 70% or more with a follow-up period of approximately 24 months after CAR-T cell infusion. In some embodiments, the method is effective in maintaining progression-free survival with a rate between approximately 79% and approximately 93% with a follow-up period of approximately 6 months after CAR-T cell infusion, a rate between approximately 67% and approximately 84% with a follow-up period of approximately 12 months after CAR-T cell infusion, and a rate between approximately 57% and approximately 75% with a follow-up period of approximately 18 months after CAR-T cell infusion.a rate between approximately 57% and approximately 75% with a follow-up period of approximately 21 months after CAR-T cell infusion, or a rate between approximately 49% and approximately 70% with a follow-up period of approximately 24 months after CAR-T cell infusion. In some embodiments, the method is effective in maintaining progression-free survival with a rate of approximately 88% with a follow-up period of approximately 6 months after CAR-T cell infusion, a rate of approximately 76% with a follow-up period of approximately 12 months after CAR-T cell infusion, a rate of approximately 67% with a follow-up period of approximately 18 months after CAR-T cell infusion, a rate of approximately 67% with a follow-up period of approximately 21 months after CAR-T cell infusion, or a rate of approximately 61% with a follow-up period of approximately 24 months after CAR-T cell infusion.

[0212] In some embodiments, the method further includes treating the subject for cytokine release syndrome for more than approximately 1 day after CAR-T cell infusion. In some embodiments, the method is effective in achieving a recovery rate from cytokine release syndrome of between approximately 1% and approximately 99% at a time of approximately 1, 3, 4, 6, 16, or 97 days after the first observation of cytokine release syndrome.

[0213] In some embodiments, the method further comprises treating the subject for immune effector cell-associated neurotoxicity for more than approximately 3 days after CAR-T cell infusion. In some embodiments, the method is effective in obtaining a recovery rate from immune effector cell-associated neurotoxicity of between approximately 1% and approximately 17% at a time of approximately 1, 4, 5, 8, 12, or 16 days after the first observation of immune effector cell-associated neurotoxicity.

[0214] In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in the subject. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 90% of subjects. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 91% of subjects. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 92% of subjects. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 93% of subjects. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 94% of subjects. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 95% of subjects.In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 96% of subjects. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 97% of subjects. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 98% of subjects. In certain embodiments, the treatment method is effective in maintaining a reduced tumor burden in more than 99% of subjects. In some embodiments, the treatment method is effective in maintaining a reduced tumor burden in 100% of subjects.

[0215] In certain embodiments, the treatment method is effective in maintaining a negative minimal residual disease (MRD) status in the subject. In certain embodiments, the treatment method is effective in maintaining a negative minimal residual disease (MRD) status in the subject. In certain embodiments, the treatment method is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -6 in the subject. In certain embodiments, the treatment procedure is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -5 in the subject. In certain embodiments, the treatment procedure is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -4in the subject. In certain embodiments, the treatment procedure is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -3in the subject. In certain embodiments, the treatment procedure is effective in maintaining an MRD-negative status when evaluated in bone marrow. In certain embodiments, the treatment procedure is effective in maintaining an MRD-negative status when evaluated using a bone marrow sample that is evaluable. In certain embodiments, the treatment procedure is effective in maintaining an MRD-negative status when evaluated using bone marrow DNA. In certain embodiments, the treatment procedure is effective in maintaining an MRD-negative status when evaluated at a follow-up period of more than or equal to 28 days after CAR-T cell administration. In certain embodiments, the treatment procedure is effective in maintaining an MRD-negative status when evaluated at a follow-up period of more than or equal to 1 month after CAR-T cell administration.In certain embodiments, the treatment method is effective in maintaining an MRD-negative status when evaluated at a follow-up period of 3 months or more after CAR-T cell administration. In certain embodiments, the treatment method is effective in maintaining an MRD-negative status when evaluated at a follow-up period of 6 months or more after CAR-T cell administration. In certain embodiments, the treatment method is effective in maintaining an MRD-negative status when evaluated at a follow-up period of 9 months or more after CAR-T cell administration. In certain embodiments, the treatment method is effective in maintaining an MRD-negative status when evaluated at a follow-up period of 12 months or more after CAR-T cell administration.

[0216] In certain embodiments, the treatment method is effective in maintaining an initial minimal residual disease (MRD) negative status in the subject. In certain embodiments, the treatment method is effective in maintaining the MRD-negative status at a sensitivity level of 10. -5 In certain embodiments, the treatment method is effective in maintaining a negative minimal residual disease (MRD) status at a sensitivity level of 10. -6 in the subject. In certain embodiments, the treatment procedure is effective in maintaining the MRD-negative status at a sensitivity level of 10. -4 In certain embodiments, the treatment method is effective in maintaining the MRD-negative status at a sensitivity level of 10 -3In certain embodiments, the treatment procedure is effective in maintaining MRD-negative status when evaluated using a bone marrow sample. In certain embodiments, the treatment procedure is effective in maintaining MRD-negative status when evaluated using an evaluable bone marrow sample. In certain embodiments, the treatment procedure is effective in maintaining MRD-negative status when evaluated using bone marrow DNA. In certain embodiments, the treatment procedure is effective in maintaining MRD-negative status when evaluated at a follow-up period of more than or equal to 1 month after CAR-T cell administration.In certain embodiments, the treatment method is effective in maintaining MRD-negative status when evaluated at a follow-up period of 3 months or more after CAR-T cell administration. In certain embodiments, the treatment method is effective in maintaining MRD-negative status when evaluated at a follow-up period of 6 months or more after CAR-T cell administration. In certain embodiments, the treatment method is effective in maintaining MRD-negative status when evaluated at a follow-up period of 9 months or more after CAR-T cell administration. In certain embodiments, the treatment method is effective in maintaining MRD-negative status when evaluated at a follow-up period of 12 months or more after CAR-T cell administration.

[0217] In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status. In certain embodiments, the efficacy of the treatment procedure is assessed by assessing the proportion of subjects with MRD-negative status at a sensitivity level of 10. -6 In certain embodiments, the effectiveness of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status at a sensitivity level of 10. -5 In certain embodiments, the effectiveness of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status at a sensitivity level of 10. -4 In certain embodiments, the effectiveness of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status at a sensitivity level of 10. -3The efficacy of the treatment procedure is evaluated in certain embodiments by assessing the proportion of subjects with MRD-negative status at a median follow-up of 28 days or more after CAR-T cell administration. In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of individuals with MRD-negative status at a median follow-up of 1 month or more after CAR-T cell administration. In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of subjects with MRD-negative status at a median follow-up of 3 months or more after CAR-T cell administration.In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with MRD-negative status at a median follow-up of 6 months or more after CAR-T cell administration. In certain embodiments, the efficacy of the treatment procedure is assessed by assessing the proportion of subjects with MRD-negative status at a median follow-up of 9 months or more after CAR-T cell administration. In certain embodiments, the efficacy of the treatment procedure is assessed by assessing the proportion of subjects with MRD-negative status at a median follow-up of 12 months or more after CAR-T cell administration.

[0218] In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of subjects with evaluable bone marrow and MRD-negative status. In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of subjects with evaluable bone marrow and MRD-negative status at a sensitivity level of 10. -6 In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a sensitivity level of 10. -5 In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a sensitivity level of 10. -4In certain embodiments, the efficacy of the treatment procedure is evaluated by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a sensitivity level of 10. - 3The efficacy of the treatment procedure is evaluated in certain embodiments by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a median follow-up of 28 days or more after CAR-T cell administration. In certain embodiments, the efficacy of the treatment procedure is assessed by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a median follow-up of 1 month or more after CAR-T cell administration. In certain embodiments, the efficacy of the treatment procedure is assessed by assessing the proportion of subjects with evaluable bone marrow and MRD-negative status at a median follow-up of 3 months or more after CAR-T cell administration.In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of subjects with evaluable bone marrow and MRD-negative status at a median follow-up of 6 months or more after CAR-T cell administration. In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of subjects with evaluable bone marrow and MRD-negative status at a median follow-up of 9 months or more after CAR-T cell administration. In certain embodiments, the efficacy of the treatment procedure is assessed by evaluating the proportion of subjects with evaluable bone marrow and MRD-negative status at a median follow-up of 12 months or more after CAR-T cell administration.

[0219] In certain embodiments, the effectiveness of the treatment procedure is evaluated by assessing the proportion of subjects with a stringent-complete response. In certain embodiments, the effectiveness of the treatment procedure is assessed by assessing the proportion of subjects with a complete response or better. In certain embodiments, the effectiveness of the treatment procedure is assessed by assessing the proportion of subjects with a very good partial response or better. In certain embodiments, the effectiveness of the treatment procedure is assessed by assessing the proportion of subjects with a partial response or better. In certain embodiments, the effectiveness of the treatment procedure is assessed using an overall response rate. In some embodiments, the overall response rate is the proportion of subjects with a partial response or better.

[0220] In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 39% at a sensitivity threshold of 10. -5 In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 44% at a sensitivity threshold of 10. -5 In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 49% at a sensitivity threshold of 10. -5 In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 54% at a sensitivity threshold of 10. -5 In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 59% at a sensitivity threshold of 10.-5 In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 64% at a sensitivity threshold of 10. -5 In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 69% at a sensitivity threshold of 10. -5 In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 74% at a sensitivity threshold of 10. -5 In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 70% at a sensitivity threshold of 10. -5In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 75% at a sensitivity threshold of 10. -5 in evaluable bone marrow. In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 80% at a sensitivity threshold of 10. -5 in evaluable bone marrow. In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 85% at a sensitivity threshold of 10. -5 in evaluable bone marrow. In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 90% at a sensitivity threshold of 10. -5in evaluable bone marrow. In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of more than 95% at a sensitivity threshold of 10. -5 in evaluable bone marrow. In certain embodiments, the method is effective in maintaining a minimal residual disease (MRD) negativity rate of 100% at a sensitivity threshold of 10. -5 in the evaluable bone marrow.

[0221] In certain embodiments, the treatment method is effective in obtaining an overall reaction rate of more than 75%. In certain embodiments, the treatment method is effective in obtaining an overall reaction rate of more than 80%. In certain embodiments, the treatment method is effective in obtaining an overall reaction rate of more than 85%. In certain embodiments, the treatment method is effective in obtaining an overall reaction rate of more than 90%. In certain embodiments, the treatment method is effective in obtaining an overall reaction rate of more than 91%. In certain embodiments, the method is effective in obtaining an overall reaction rate of more than 93%. In certain embodiments, the method is effective in obtaining an overall reaction rate of more than 95%. In certain embodiments, the method is effective in obtaining an overall reaction rate of more than 97%.In certain embodiments, the method is effective in obtaining an overall response rate of more than 99%. In some embodiments, the method is effective in obtaining an overall response rate of 100%. In certain embodiments, the treatment method is effective in obtaining an overall response rate of approximately 84.6%. In certain embodiments, the treatment method is effective in obtaining an overall response rate of approximately 99.4%. In certain embodiments, the overall response rate is evaluated at a median follow-up period of at least 6 months after CAR-T cell infusion. In certain embodiments, the overall response rate is evaluated at a median follow-up period of at least 12 months after CAR-T cell infusion.

[0222] In certain embodiments, more than 70% of subjects respond to the treatment 9 months after CAR-T cell administration. In certain embodiments, more than 72% of subjects respond to the treatment 9 months after CAR-T cell administration. In certain embodiments, more than 74% of subjects respond to the treatment 9 months after CAR-T cell administration. In certain embodiments, more than 76% of subjects respond to the treatment 9 months after CAR-T cell administration. In certain embodiments, more than 78% of subjects respond to the treatment 9 months after CAR-T cell administration. In certain embodiments, more than 80% of subjects respond to the treatment 9 months after CAR-T cell administration.In certain embodiments, more than 82% of subjects respond to the treatment 9 months after CAR-T cell administration. In certain embodiments, more than 84% of subjects respond to the treatment 9 months after CAR-T cell administration. In certain embodiments, more than 86% of subjects respond to the treatment 9 months after CAR-T cell administration.

[0223] In certain embodiments, more than 54% of responding subjects respond to the treatment 12 months after CAR-T cell administration. In certain embodiments, more than 58% of responding subjects respond to the treatment 12 months after CAR-T cell administration. In certain embodiments, more than 62% of responding subjects respond to the treatment 12 months after CAR-T cell administration. In certain embodiments, more than 66% of responding subjects respond to the treatment 12 months after CAR-T cell administration. In certain embodiments, more than 70% of responding subjects respond to the treatment 12 months after CAR-T cell administration. In certain embodiments, more than 74% of responding subjects respond to the treatment 12 months after CAR-T cell administration.In certain formulations, more than 78% of responding subjects respond to the treatment procedure 12 months after administration of CAR-T cells.

[0224] In certain embodiments, the treatment method is effective in maintaining a reaction time of more than 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 months or longer. In certain embodiments, the treatment method is effective in maintaining a reaction time of more than 12.4 months. In certain embodiments, the treatment method is effective in maintaining a reaction time of more than 15.9 months.

[0225] In certain embodiments, the treatment method is effective in maintaining a median response time of more than 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 months or longer. In certain embodiments, the treatment method is effective in maintaining a median response time of more than 12.4 months. In certain embodiments, the treatment method is effective in maintaining a median response time of more than 15.9 months.

[0226] In certain embodiments, the treatment method is effective in obtaining a complete response or better in more than 60% of the subjects. In certain embodiments, the treatment method is effective in obtaining a complete response or better in more than 61% of the subjects. In certain embodiments, the treatment method is effective in obtaining a complete response or better in more than 62% of the subjects. In certain embodiments, the treatment method is effective in obtaining a complete response or better in more than 63% of the subjects. In certain embodiments, the treatment method is effective in obtaining a complete response or better in more than 64% of the subjects. In certain embodiments, the treatment method is effective in obtaining a complete response or better in more than 65% of the subjects.In certain embodiments, the treatment method is effective in achieving a complete response or better in more than 66% of subjects. In certain embodiments, the treatment method is effective in achieving a complete response or better in more than 67% of subjects. In certain embodiments, the complete response or better is assessed less than 1 month after CAR-T cell administration. In certain embodiments, the complete response or better is assessed less than 3 months after CAR-T cell administration. In certain embodiments, the complete response or better is assessed less than 6 months after CAR-T cell administration. In certain embodiments, the complete response or better is assessed less than 9 months after CAR-T cell administration.In certain embodiments, the complete response or better is evaluated less than 12 months after CAR-T cell administration. In certain embodiments, the complete response or better is evaluated less than 15 months after CAR-T cell administration. In certain embodiments, the complete response or better is evaluated more than 15 months after CAR-T cell administration.

[0227] In certain embodiments, the treatment method is effective in obtaining a very good partial response or better in more than 80% of the subjects. In certain embodiments, the treatment method is effective in obtaining a very good partial response or better in more than 85% of the subjects. In certain embodiments, the treatment method is effective in obtaining a very good partial response or better in more than 86% of the subjects. In certain embodiments, the treatment method is effective in obtaining a very good partial response or better in more than 87% of the subjects. In certain embodiments, the treatment method is effective in obtaining a very good partial response or better in more than 88% of the subjects. In certain embodiments, the treatment method is effective in obtaining a very good partial response or better in more than 89% of the subjects.In certain embodiments, the treatment method is effective in achieving a very good partial response or better in more than 90% of subjects. In certain embodiments, the treatment method is effective in achieving a very good partial response or better in more than 91% of subjects. In certain embodiments, the treatment method is effective in achieving a very good partial response or better in more than 92% of subjects. In certain embodiments, the very good partial response or better is assessed less than 1 month after CAR-T cell administration. In certain embodiments, the very good partial response or better is assessed less than 3 months after CAR-T cell administration. In certain embodiments, the very good partial response or better is assessed less than 6 months after CAR-T cell administration.In certain embodiments, a very good partial response or better is evaluated less than 9 months after CAR-T cell administration. In certain embodiments, a very good partial response or better is evaluated less than 12 months after CAR-T cell administration. In certain embodiments, a very good partial response or better is evaluated less than 15 months after CAR-T cell administration. In certain embodiments, a very good partial response or better is evaluated more than 15 months after CAR-T cell administration.

[0228] In certain embodiments, the treatment method is effective in maintaining a median time to first response of less than 1.15 months. In certain embodiments, the treatment method is effective in maintaining a median time to first response of less than 1.10 months. In certain embodiments, the treatment method is effective in maintaining a median time to first response of less than 1.05 months. In certain embodiments, the treatment method is effective in maintaining a median time to first response of less than 1.00 months. In certain embodiments, the treatment method is effective in maintaining a median time to first response of less than 0.95 months.

[0229] In certain embodiments, the treatment method is effective in maintaining a median time to best response of less than 2.96 months. In certain embodiments, the treatment method is effective in maintaining a median time to best response of less than 2.86 months. In certain embodiments, the treatment method is effective in maintaining a median time to best response of less than 2.76 months. In certain embodiments, the treatment method is effective in maintaining a median time to best response of less than 2.66 months. In certain embodiments, the treatment method is effective in maintaining a median time to best response of less than 2.56 months.

[0230] In certain embodiments, the method is effective in maintaining an overall survival rate of more than 80% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 82% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 85% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 87% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 90% after 9 months following CAR-T cell administration.In certain embodiments, the method is effective in maintaining an overall survival rate of more than 92% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 95% after 9 months following CAR-T cell administration.

[0231] In certain embodiments, the method is effective in maintaining an overall survival rate of more than 80% after 12 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 83% after 12 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 86% after 12 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 89% after 12 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining an overall survival rate of more than 92% after 12 months following CAR-T cell administration.In certain embodiments, the method is effective in maintaining an overall survival rate of more than 93% after 12 months following CAR-T cell administration.

[0232] In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 70% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 72% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 75% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 77% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 80% after 9 months following CAR-T cell administration.In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 82% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 85% after 9 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than or equal to 87% after 9 months following CAR-T cell administration.

[0233] In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 66% after 12 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 69% after 12 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 72% after 12 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 76% after 12 months following CAR-T cell administration. In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 80% after 12 months following CAR-T cell administration.In certain embodiments, the method is effective in maintaining a progression-free survival rate of more than 84% after 12 months following CAR-T cell administration.

[0234] In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 86% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 88% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 90% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 92% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 94% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 96% of subjects.In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 98% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 99% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from cytokine release syndrome in more than 100% of subjects.

[0235] In certain embodiments, the treatment method is effective in maintaining recovery from immune effector cell-associated neurotoxicity, if present, in more than 90% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from immune effector cell-associated neurotoxicity, if present, in more than 92% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from immune effector cell-associated neurotoxicity, if present, in more than 94% of subjects. In certain embodiments, the treatment method is effective in maintaining recovery from immune effector cell-associated neurotoxicity, if present, in more than 96% of subjects.In certain embodiments, the treatment method is effective in achieving recovery from immune effector cell-associated neurotoxicity, if present, in more than 98% of subjects. In certain embodiments, the treatment method is effective in achieving recovery from immune effector cell-associated neurotoxicity, if present, in more than 100% of subjects.

[0236] In some embodiments, the method further includes diagnosing the subject for cytopenias. In some embodiments, the cytopenias include one, several, or all of lymphopenia, neutropenia, and thrombocytopenia. Without being bound to any theory, grade 3 or 4 lymphopenia, but not grade 2 or lower, is defined by a lymphocyte count of less than 0.5 × 10⁻⁵. 9Grade 3 or 4 neutropenia, but not grade 2 or lower, is characterized by a neutrophil count of less than 1,000 cells per microliter of a subject's blood sample, and grade 3 or 4 thrombocytopenia, but not grade 2 or lower, is characterized by a platelet count of less than 50,000 cells per microliter of the subject's blood sample. In some embodiments, more than 75% of subjects with grade 3 or 4 lymphopenia recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 80% of subjects with grade 3 or 4 lymphopenia recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration.In some embodiments, more than 85% of subjects with grade 3 or 4 lymphopenia recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 90% of subjects with grade 3 or 4 lymphopenia recover to grade 2 or lower lymphopenia 60 days after CAR-T cell administration. In some embodiments, more than 70% of subjects with grade 3 or 4 neutropenia recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 75% of subjects with grade 3 or 4 neutropenia recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration.In some embodiments, more than 80% of subjects with grade 3 or 4 neutropenia recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 85% of subjects with grade 3 or 4 neutropenia recover to grade 2 or lower neutropenia 60 days after CAR-T cell administration. In some embodiments, more than 30% of subjects with grade 3 or grade 4 thrombocytopenia recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration. In some embodiments, more than 34% of subjects with grade 3 or grade 4 thrombocytopenia recover to grade 2 or lower thrombocytopenia 60 days after CAR-T cell administration.In some embodiments, more than 38% of subjects with grade 3 or grade 4 thrombocytopenia recover to grade 2 or lower 60 days after CAR-T cell administration. In some embodiments, more than 42% of subjects with grade 3 or grade 4 thrombocytopenia recover to grade 2 or lower 60 days after CAR-T cell administration.

[0237] In certain embodiments, the subject is retreated by administering a second intravenous infusion of a second dose of CAR-T cells. In certain embodiments, the retreatment dose comprises 1.0 × 10 5 up to 5.0 × 10 6 CAR-T cells per kilogram of the subject's mass. In certain embodiments, the post-treatment dose comprises approximately 0.75 × 10 5CAR-T cells per kilogram of the subject's mass. In certain embodiments, the subject is retreated after the onset of disease progression, following a best response of minimal response or better after the first CAR-T cell infusion. In certain embodiments, the time between the first CAR-T cell infusion and the detection of disease progression is at least six months. Kits and manufacturing materials

[0238] Any of the compositions described here can be included in a kit. In some embodiments, engineered immortalized CAR-T cells are provided in the kit, which may also include reagents suitable for expanding the cells, such as media.

[0239] In a non-restrictive example, a chimeric receptor expression construct, one or more reagents for generating a chimeric receptor expression construct, cells for transfecting the expression construct and / or one or more instruments for obtaining immortalized T cells for transfection of the expression construct (such an instrument may be a syringe, a pipette, tweezers and / or other medically approved device).

[0240] In some embodiments, the kit includes reagents or devices for electroporation of cells.

[0241] In some embodiments, the kit includes artificial antigen-presenting cells.

[0242] The kits may comprise one or more suitably aliquoted compositions of the present disclosure or reagents for generating compositions of the disclosure. The components of the kits may be packaged either in aqueous medium or in lyophilized form. The containers of the kits may include at least one vial, test tube, flask, bottle, syringe, or other container into which a component can be placed and preferably aliquoted in a suitable manner. If more than one component is present in the kit, the kit generally also includes a second, third, or other additional container into which the additional components can be placed separately. However, different combinations of components may be contained in a single ampoule.The kits described in this disclosure typically include a means for storing the chimeric receptor construct and all other reagent containers in a tightly sealed container for commercial sale. Such containers may be injection-molded or blow-molded plastic containers, in which, for example, the desired vials are retained.

[0243] The following examples are intended to be purely illustrative of the disclosure and should therefore not be regarded as a limitation of the disclosure in any way. EXAMPLES OF EXECUTION 1. Method for treating a subject, comprising administering a dose of T cells comprising a chimeric antigen receptor (CAR) to the subject, comprising: (a) an extracellular antigen-binding domain capable of binding specifically to an epitope of the B-cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, where the subject has multiple myeloma, has received one to three prior lines of therapy, including immunomodulatory drug (IMiD) therapy, and is refractory to IMiD. 2. Method according to embodiment 1, wherein the subject has a high-risk feature and wherein, optionally, the high-risk feature is a cytogenetic abnormality, an International Staging System (ISS) stage III and / or soft tissue plasmacytomas. 3. Procedures for the selective treatment of a subject, comprising: (1) Determine whether the subject has a high-risk feature, wherein the high-risk feature is a cytogenetic abnormality, an International Staging System (ISS) stage III and / or soft tissue plasmacytomas; and (2) Administering a dose of T cells comprising a chimeric antigen receptor (CAR) to the subject determined to have the high-risk characteristic in step (1), comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of BCMA, (b) a transmembrane domain, and (c) an intracellular signaling domain, where the subject may have multiple myeloma, has received one to three prior lines of therapy, including therapy with an IMiD, and is refractory to the IMiD. 4. Method for the selective treatment of a subject, comprising administering a dose of T cells comprising a chimeric antigen receptor (CAR) to the subject in whom a high-risk trait has been identified, including: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of BCMA, (b) a transmembrane domain, and (c) an intracellular signaling domain, The high-risk feature is a cytogenetic abnormality, an International Staging System (ISS) stage III and / or soft tissue plasmacytomas, where the subject may have multiple myeloma, has received one to three prior lines of therapy, including therapy with an IMiD, and is refractory to the IMiD. 5. Method according to one of embodiments 1-4, wherein the IMiD is lenalidomide. 6. Method according to one of embodiments 2-5, wherein the high-risk feature is a cytogenetic abnormality. 7. Method according to embodiment 6, wherein the cytogenetic anomaly is a high-risk cytogenetic anomaly. 8. Method according to embodiment 7, wherein the subject has one or more high-risk cytogenetic abnormalities selected from a group comprising Gain / AMP(1q), del(17p), t(4;14), t(14;16) or any combination thereof. 9. Method according to embodiment 8, wherein the cytogenetic anomaly comprises Gain / amp(1q). 10. Method according to embodiment 8, wherein the cytogenetic anomaly comprises del(17p). 11. Method according to embodiment 8, wherein the cytogenetic anomaly comprises t(4;14). 12. Method according to embodiment 8, wherein the cytogenetic anomaly comprises t(14;16). 13. Method according to one of embodiments 6-12, wherein the subject has at least two cytogenetic abnormalities and wherein the subject optionally has two, three, four, five or more cytogenetic abnormalities. 14. Method according to embodiment 6, wherein the cytogenetic anomaly is a standard risk cytogenetic anomaly. 15. Method according to one of embodiments 2-5, wherein the high-risk feature is International Staging System (ISS) Level III. 16. Method according to one of embodiments 2-5, wherein the high-risk feature is soft tissue plasmacytomas. 17. Method according to one of embodiments 1-16, wherein the subject has received a prior line of therapy. 18. Method according to one of embodiments 1-16, wherein the subject has received two prior lines of therapy. 19. Method according to one of embodiments 1-16, wherein the subject has received three prior lines of therapy. 20. Method according to one of embodiments 1-19, wherein the one, two or three prior lines of therapy include treatment with pomalidomide. 21. Method according to any embodiment 1-20, wherein the one, two or three prior lines of therapy further comprise treatment with an anti-CD38 antibody and wherein the anti-CD38 antibody is optionally daratumumab and / or isatuximab. 22. Method according to any embodiment 1-21, wherein the one, two or three prior lines of therapy further comprise treatment with a proteasome inhibitor, wherein the proteasome inhibitor is optionally bortezomib, carfilzomib, ixazomib or any combination thereof. 23. Method according to any one of embodiments 1-22, wherein the subject has further received bridge therapy, wherein the bridge therapy may optionally be of the physician's choice, wherein the bridge therapy may optionally include pomalidomide, bortezomib, dexamethasone, daratumumab or any combination thereof, wherein the bridge therapy may further optionally include pomalidomide, bortezomib and dexamethasone, and wherein the bridge therapy may further optionally include daratumumab, pomalidomide and dexamethasone. 24. Method according to embodiment 23, wherein the subject received the bridge therapy from about every 20 days to about every 30 days, wherein the subject optionally received the bridge therapy about every 21 days, wherein the subject optionally received the bridge therapy about every 28 days, and wherein furthermore, optionally, the subject received at least one, two, three, four or more bridge therapies. 25. Method according to one of embodiments 1-24, wherein the subject has additionally received lymphodepletion therapy, wherein the lymphodepletion therapy optionally comprises cyclophosphamide and / or fludarabine daily, wherein the lymphodepletion therapy optionally comprises cyclophosphamide and fludarabine daily, wherein the lymphodepletion treatment optionally comprises cyclophosphamide at a concentration of about 300 mg / m² 2 and fludarabine at a concentration of approximately 30 mg / m³ 2 daily over 3 days. 26. Method according to one of embodiments 1-25, wherein the dose of the T cells is 0.5-1.0 × 10 6 -cells / kg body weight of the subject, where, if necessary, the dose of T cells is approximately 0.75 × 10 6-cells / kg body weight of the subject, the procedure possibly comprising administering the dose of T cells approximately 5 to approximately 7 days after the start of lymphodepletion therapy, the dose possibly being administered as a single infusion. 27. Method according to any one of embodiments 1-26, wherein the method is effective in maintaining an overall response in the subject after administration of the dose of T cells to the subject, wherein optionally the method is effective in maintaining the overall response at a rate of about 75% to about 100%, wherein further optionally the method is effective in maintaining the overall response at a rate of about 84.6%, and wherein further optionally the method is effective in maintaining the overall response at a rate of about 99.4%. 28. Method according to embodiment 27, wherein the overall response, in order from best to worst, comprises the following: (1) a stringent and complete response; (2) a complete reaction; (3) a very good partial response; (4) a partial reaction; or (5) a minimal response. 29. Method according to embodiment 28, wherein the overall reaction is a stringent complete reaction, wherein the method is optionally effective in obtaining the stringent complete reaction at a rate of about 40% to about 90%, about 50% to about 80%, about 58.2% or about 68.8%. 30. Method according to embodiment 28, wherein the overall reaction is a complete reaction, wherein optionally the method is effective to obtain the complete reaction at a rate of about 10% to about 20%, wherein further optionally the method is effective to obtain the complete reaction at a rate of about 14.9% or about 17.6%. 31. Method according to embodiment 28, wherein the overall reaction is a very good partial reaction or a partial reaction. 32. Method according to embodiment 28, wherein: (1) the process is effective in obtaining a stringent complete reaction or a complete reaction at a rate of about 70% to about 90%, and optionally the process is effective in obtaining a stringent complete reaction or a complete reaction at a rate of about 73.1% or about 86.4%; (2) the process is effective in obtaining a stringent complete reaction, a complete reaction or a very good partial reaction at a rate of about 80% to about 100%, wherein the process is optionally effective in obtaining a stringent complete reaction, a complete reaction or a very good partial reaction at a rate of about 81.3% or about 96.0%; (3) the procedure is effective in obtaining a minimal response; (4) the procedure is effective for maintaining a minimal residual disease negative, where the procedure may be effective for maintaining a minimal residual disease negative at a rate of approximately 50% to approximately 80%, where the procedure may continue to be effective for maintaining a minimal residual disease negative at a rate of approximately 60.6% or approximately 71.6%; or (5) the procedure shall be effective in further maintaining 12-month progression-free survival in at least about 60% to about 100% of subjects, in at least about 69.4% to about 81.1% of subjects or in at least about 84.1% to about 93.4% of subjects, where appropriate the procedure shall be effective in maintaining 12-month progression-free survival in at least about 75.9% of subjects or in about 89.7% of subjects. 33. Method according to one of embodiments 27-32, wherein: (1) the time to the first overall response or first minimal response is in the range of approximately 0.9 to approximately 11.1 months, with the median time to the first overall response or first minimal response being approximately 2.1 months; or (2) the time to best overall response or best minimum response is approximately 1.1 to approximately 18.6 months, where the median time to best overall response or best minimum response may be approximately 6.4 or approximately 6.5 months. 34. A method according to any one of embodiments 1-33, wherein the method further comprises treating the subject for an adverse event following administration of the dose of T cells, wherein the method optionally comprises administering treatment to the subject to alleviate the adverse event, wherein optionally the adverse event comprises a hematological adverse event, a non-hematological adverse event, a treatment-related adverse event, or any combination thereof, wherein optionally the non-hematological adverse event comprises an infection other than an infection and / or a non-hematological adverse event other than an infection, wherein optionally the adverse event comprises neutropenia, thrombocytopenia, anemia, lymphopenia, an upper respiratory tract infection, nasopharyngitis, sinusitis, rhinitis, tonsillitis, pharyngitis, laryngitis, pharyngotonsillitis, COVID-19,COVID-19 pneumonia, asymptomatic COVID-19, neutropenic sepsis, progressive multifocal leukoencephalopathy, septic shock, respiratory failure, pulmonary embolism, lower respiratory tract / lung infection, pneumonia, bronchitis, nausea, hypogammaglobulinemia, diarrhea, fatigue, headache, constipation, hypokalemia, asthenia, peripheral edema, decreased appetite, peripheral sensory neuropathy, back pain, arthralgia, pyrexia, dyspnea, insomnia, or any combination thereof, where the adverse event is, where appropriate, a Grade 3 / 4 adverse event and where, where appropriate, the adverse event lasts for more than approximately 30 days or approximately 60 days. 35.A method according to any one of embodiments 1-34, wherein the method further comprises treating the subject for a second primary malignancy after administering the dose of T cells, wherein the method optionally comprises administering a treatment to the subject to alleviate the second primary malignancy, wherein optionally the second primary malignancy comprises a cutaneous / non-invasive malignancy, a hematological malignancy, a non-cutaneous / invasive malignancy, or any combination thereof, wherein optionally the second primary malignancy comprises basal cell carcinoma, Bowen's disease, lip squamous cell carcinoma, malignant melanoma, malignant melanoma in situ, cutaneous squamous cell carcinoma, acute myeloid leukemia, a myelodysplastic syndrome, peripheral T-cell lymphoma, angiosarcoma, invasive lobular breast carcinoma, pleomorphic malignant fibrous histiocytoma, renal cell carcinoma, Tonsil carcinoma or any combination thereof. 36. Method according to embodiment 34 or 35, wherein the adverse event or second primary malignancy occurs in the subject at a rate comparable to the rate of the same adverse event or second primary malignancy occurring in a subject undergoing standard treatment. 37. A method according to any one of embodiments 1-36, wherein the method further comprises treating the subject against a CAR-T-associated adverse event after administration of the dose of T cells, wherein the method optionally comprises administering treatment to the subject to alleviate the CAR-T-associated adverse event, wherein optionally the CAR-T-associated adverse event comprises cytokine release syndrome (CRS) and / or neurotoxicity, wherein optionally: (a) the CAR-T-associated adverse event is a CRS, wherein the CRS may further occur in the subject at a rate of approximately 60% to approximately 90% or at a rate of approximately 76.1%, wherein the maximum toxicity grade of the CRS may further be Grade 1, Grade 2 or Grade 3, wherein further: (1) the maximum toxicity level of the CRS is Grade 1, where, where applicable, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 52.8%; (2) the maximum toxicity level of the CRS is grade 2, where, where applicable, the maximum toxicity level of grade 2 occurs in the subject at a rate of approximately 22.2%; (3) the maximum toxicity level of the CRS is grade 3, where, where applicable, the maximum toxicity level of grade 3 occurs in the subject at a rate of approximately 1.1%; (4) the time to first onset of CRS is in the range of about 1 to about 23 days, with a median time to first onset of CRS of about 8 days where applicable; (5) the duration of the CRS ranges from about 1 to about 17 days, with a median duration of about 3 days where applicable; or (6) the treatment includes tocilizumab, oxygen, a corticosteroid, a vasopressor or any combination thereof; or (b) the CAR-T-associated adverse event is neurotoxicity, where appropriate the neurotoxicity includes an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, standard treatment-arm adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, where appropriate the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated neurotoxicity: (1) The immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of approximately 4.5%; (2) the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1 or Grade 2, wherein the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1, furthermore, where appropriate, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 3.4%, or where, where appropriate, the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 2, furthermore, where appropriate, the maximum toxicity level of Grade 2 occurs in the subject at a rate of approximately 1.1%; (3) the time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 6 to about 15 days, with a median time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom of about 9.5 days, if applicable; (4) the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 1 to about 6 days, with the median duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom being about 2 days; or (5) the treatment includes a corticosteroid and / or tocilizumab. 38. Method according to embodiment 37, wherein the neurotoxicity is CAR-T cell neurotoxicity, wherein optionally the CAR-T cell neurotoxicity occurs in the subject at a rate of about 17.0%, wherein optionally the CAR-T cell neurotoxicity comprises Grade 3 / 4 neurotoxicity, Grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, an adverse event occurring with exercise and neurocognitive treatment, or any combination thereof, wherein further optionally: (a) CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, occurring in approximately 2.3% of the subject; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, with cranial nerve palsy occurring in the subject at a rate of approximately 9.1%, where: (1) the cranial nerve palsy is grade 2 or grade 3, where furthermore, where applicable, the cranial nerve palsy is grade 2, which occurs in the subject at a rate of about 8.0%, and where furthermore, where applicable, the cranial nerve palsy is grade 3, which occurs in the subject at a rate of about 1.1%, (2) the time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject ranges from about 17 days to about 60 days, and furthermore, where applicable, the median time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject is about 21 days; (3) the cranial nerve palsy affects cranial nerve III, V or VII; (4) the duration of cranial nerve palsy ranges from about 15 days to about 262 days, with the median duration of cranial nerve palsy being about 77 days; or (5) the treatment includes a corticosteroid; (d) CAR T-cell neurotoxicity is a peripheral neuropathy, with peripheral neuropathy occurring in the subject at a rate of approximately 2.8%, if applicable; or (e) CAR-T cell neurotoxicity is an adverse event induced by exercise and neurocognitive treatment, wherein, where appropriate, the adverse event induced by exercise and neurocognitive treatment is grade 1, and furthermore, where, where appropriate, the grade 1 adverse event induced by neurocognitive treatment and the subject's adverse event induced by neurocognitive treatment occur at a rate of approximately 0.6%. 39. Method according to one of embodiments 1-38, wherein: (a) CD3+ cells encompassing the CAR in the subject's blood reach their median peak approximately 13 days after administration of the T cells to the subject, with the CD3+ cells encompassing the CAR in the subject's blood potentially reaching a peak at a mean concentration of approximately 1523 cells / µl; (b) CD3+ cells encompassing the CAR remain detectable in the subject's blood from approximately 13 days to approximately 631 days after administration of the T cells to the subject, with the CD3+ cells encompassing the CAR remaining detectable in the subject's blood for a median of approximately 57 days after administration of the T cells to the subject, if applicable; or (c) the AUC 0-28 The average number of CD3+ cells encompassing the CAR in the subject's blood is approximately 12,504 cells / µL. 40. A method according to one of embodiments 1-39, wherein the first VHH domain comprises a CDR1, a CDR2, and a CDR3 as shown in the VHH domain, comprising the amino acid sequence of SEQ ID NO: 2, and the second VHH domain comprises a CDR1, a CDR2, and a CDR3 as shown in the VHH domain, comprising the amino acid sequence of SEQ ID NO: 4, wherein optionally the first VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 18, a CDR2 comprising the amino acid sequence of SEQ ID NO: 19, a CDR3 comprising the amino acid sequence of SEQ ID NO: 20, and the second VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a CDR2 comprising the amino acid sequence of SEQ ID NO: 22, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 23, wherein optionally the first VHH domain the amino acid sequence of SEQ ID NO: 2 and the second VHH domain comprises the amino acid sequence of SEQ ID NO: 4,where the first VHH domain may be located at the N-terminus of the second VHH domain, or where the first VHH domain may be located at the C-terminus of the second VHH domain, furthermore where: (a) the first VHH domain is linked to the second VHH domain via a linker comprising the amino acid sequence of SEQ ID NO: 3; (b) the transmembrane domain is derived from a molecule selected from the group consisting of CD8α, CD4, CD28, CD137, CD80, CD86, CD152 and PD1, optionally the transmembrane domain being derived from CD8α and the amino acid sequence being SEQ ID NO: 6; (c) the intracellular signaling domain comprises a primary intracellular signaling domain of an immune effector cell, the primary intracellular signaling domain optionally being derived from CD3ζ, comprising the amino acid sequence of SEQ ID NO: 8; (d) the intracellular signaling domain comprises a costimulatory signaling domain, wherein optionally the costimulatory signaling domain is derived from a costimulatory molecule selected from the group consisting of CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, ligands of CD83 and any combination thereof, wherein optionally the costimulatory signaling domain comprises a cytoplasmic domain of CD137 comprising the amino acid sequence of SEQ ID NO: 7; (e) the CAR further comprises a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain, the hinge domain optionally being derived from CD8α, comprising the amino acid sequence of SEQ ID NO: 5; (f) the CAR further comprises a signal peptide located at the N-terminus of the polypeptide, the signal peptide optionally being derived from CD8α, comprising the amino acid sequence of SEQ ID NO: 1; or (g) of the CAR comprises the amino acid sequence of SEQ ID NO: 17. 41. Method according to one of embodiments 1-40, wherein the dose of T cells is formulated in a composition comprising 5% dimethyl sulfoxide (DMSO). 42. Method according to one of embodiments 1-41, wherein the administration of the dose of T cells reduces the risk of disease progression or death in the subject. 43. Method according to embodiment 42, wherein the risk of disease progression or death is reduced compared to treatment with daratumumab pomalidomide dexamethasone (DPd) or pomalidomide bortezomib dexamethasone (PVd). 44. Method according to embodiment 42, wherein the risk of disease progression or death is reduced compared to administering ide-cell treatment. 45. Method according to one of embodiments 42-44, wherein the subject has an approximately 60% to approximately 75% reduced risk of disease progression or death. 46. ​​Method of embodiment 45, wherein the subject has an approximately 74% reduced risk of disease progression or death. 47. Method according to one of embodiments 42-46, wherein the IMiD is lenalidomide. 48. Method according to one of embodiments 42-47, wherein the subject has received three or fewer prior lines of therapy. 49. Method according to one of embodiments 42-47, wherein the subject has received two or fewer prior lines of therapy. 50. Method according to one of embodiments 42-47, wherein the subject has received only one prior line of therapy. 51. Method according to one of embodiments 42-50, wherein the method is effective in obtaining an overall reaction rate (ORR) of about 75% to about 100%. 52. Method according to embodiment 51, wherein the ORR is approximately 84.6%. 53. Method according to one of embodiments 42-52, wherein the treatment is effective in maintaining the median progression-free survival (PFS) of the subject compared to the administration of DPd or PVd treatment. 54. Method according to embodiment 53, wherein the PFS after 12 months following administration of the treatment is approximately 75.9%. 55. Method according to embodiment 54, wherein the PFS at 12 months after administration of DPd or PVd is approximately 48.6%. 56. Method according to one of embodiments 1-23 and 42-55, wherein the treatment is more effective in obtaining a stringent complete response (sCR) in the subject compared to the administration of a DPd or PVd treatment. 57. Method according to embodiment 56, wherein the sCR after administration of the treatment is approximately 58.2%. 58. Method according to embodiment 57, wherein the sCR after administration of DPd or PVd is about 15.2%. 59. Method according to one of embodiments 1-23 and 42-58, wherein the treatment is more effective in obtaining a very good partial response (VGPR) or better in the subject compared to the administration of a DPd or PVd treatment. 60. Method according to embodiment 59, wherein the VGPR or better response after administration of the treatment is about 81.3%. 61. Method according to embodiment 60, wherein the VGPR or better response after administration of DPd or PVd is about 45.5%. 62. A method according to any one of embodiments 42-61, wherein the method further comprises treating the subject against a CAR-T-associated adverse event after administration of the dose of T cells, wherein the method optionally comprises administering a treatment to the subject to alleviate the CAR-T-associated adverse event, wherein optionally the CAR-T-associated adverse event comprises cytokine release syndrome (CRS) and / or neurotoxicity, wherein optionally: (a) the CAR-T-associated adverse event is a CRS, wherein the CRS may further occur in the subject at a rate of approximately 60% to approximately 90% or at a rate of approximately 76.1%, wherein the maximum toxicity grade of the CRS may further be Grade 1, Grade 2 or Grade 3, wherein further: (1) the maximum toxicity level of the CRS is Grade 1, where, where applicable, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 52.8%; (2) the maximum toxicity level of the CRS is grade 2, where, where applicable, the maximum toxicity level of grade 2 occurs in the subject at a rate of approximately 22.2%; (3) the maximum toxicity level of the CRS is grade 3, where, where applicable, the maximum toxicity level of grade 3 occurs in the subject at a rate of approximately 1.1%; (4) the time to first onset of CRS is in the range of about 1 to about 23 days, with a median time to first onset of CRS of about 8 days where applicable; (5) the duration of the CRS ranges from about 1 to about 17 days, with a median duration of about 3 days where applicable; or (6) the treatment includes tocilizumab, oxygen, a corticosteroid, a vasopressor or any combination thereof; or (b) the CAR-T-associated adverse event is neurotoxicity, where appropriate the neurotoxicity includes an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, standard treatment-arm adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, where appropriate the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated neurotoxicity: (1) The immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of approximately 4.5%; (2) the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1 or Grade 2, wherein the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1, furthermore, where appropriate, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 3.4%, or where, where appropriate, the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 2, furthermore, where appropriate, the maximum toxicity level of Grade 2 occurs in the subject at a rate of approximately 1.1%; (3) the time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 6 to about 15 days, with a median time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom of about 9.5 days, if applicable; (4) the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 1 to about 6 days, with the median duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom being about 2 days; or (5) the treatment includes a corticosteroid and / or tocilizumab. 63. Method according to embodiment 62, wherein the neurotoxicity is CAR-T cell neurotoxicity, wherein optionally the CAR-T cell neurotoxicity occurs in the subject at a rate of about 17.0%, wherein optionally the CAR-T cell neurotoxicity comprises Grade 3 / 4 neurotoxicity, Grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, an adverse event occurring with exercise and neurocognitive treatment, or any combination thereof, wherein further optionally: (a) CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, occurring in approximately 2.3% of the subject; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, with cranial nerve palsy occurring in the subject at a rate of approximately 9.1%, where: (1) the cranial nerve palsy is grade 2 or grade 3, where furthermore, where appropriate, the cranial nerve palsy is grade 2, which occurs in the subject at a rate of about 8.0%, and where furthermore, where appropriate, the cranial nerve palsy is grade 3, which occurs in the subject at a rate of about 1.1%; (2) the time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject ranges from about 17 days to about 60 days, and furthermore, where applicable, the median time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject is about 21 days; (3) the cranial nerve palsy affects cranial nerve III, V or VII; (4) the duration of cranial nerve palsy ranges from about 15 days to about 262 days, with the median duration of cranial nerve palsy being about 77 days; or (5) the treatment includes a corticosteroid; (d) CAR T-cell neurotoxicity is a peripheral neuropathy, with peripheral neuropathy occurring in the subject at a rate of approximately 2.8%, if applicable; or (e) CAR-T cell neurotoxicity is an adverse event induced by exercise and neurocognitive treatment, wherein, where appropriate, the adverse event induced by exercise and neurocognitive treatment is grade 1, and furthermore, where, where appropriate, the grade 1 adverse event induced by neurocognitive treatment and the subject's adverse event induced by neurocognitive treatment occur at a rate of approximately 0.6%. 64. Method according to one of embodiments 42-63, wherein the subject has one or more high-risk cytogenetic abnormalities selected from a group comprising the Gain / amp(1q), del(17p), t(4;14), t(14;16) or any combination thereof. 65. Method according to embodiment 64, wherein the subject has at least two cytogenetic abnormalities and wherein the subject optionally has two, three, four, five or more cytogenetic abnormalities. 66. Method according to embodiment 64 or 65, wherein the cytogenetic anomaly is a standard risk cytogenetic anomaly. 67. Method according to any embodiment 1-66, wherein the administration of the dose of T cells is more effective in obtaining a greater very good partial response (VGPR) or better in the subject compared to the administration of a DPd or PVd treatment. 68. Method according to embodiment 67, wherein the VGPR or better response after administration of the treatment is about 81.3%. 69. Method according to embodiment 68, wherein the VGPR or better response after administration of DPd or PVd is about 45.5%. 70. Method according to one of embodiments 67-69, wherein the treatment is more effective in obtaining a stringent complete response (sCR) in the subject compared to the administration of a DPd or PVd treatment. 71. Method according to embodiment 70, wherein the sCR after administration of the treatment is approximately 58.2%. 72. Method according to embodiment 71, wherein the sCR after administration of DPd or PVd is about 15.2%. 73. Method according to one of embodiments 67-72, wherein the administration of the dose of T cells reduces the risk of disease progression or death in the subject. 74. Method according to embodiment 73, wherein the risk of disease progression or death is reduced compared to treatment with daratumumab pomalidomide dexamethasone (DPd) or pomalidomide bortezomib dexamethasone (PVd). 75. Method according to embodiment 73, wherein the risk of disease progression or death is reduced compared to administering ide-cell treatment. 76. Method according to one of embodiments 67-75, wherein the subject has a reduced risk of disease progression or death of approximately 60% to approximately 75%. 77. Method according to embodiment 76, wherein the subject has an approximately 74% reduced risk of disease progression or death. 78. Method according to one of embodiments 67-77, wherein the IMiD is lenalidomide. 79. Method according to one of embodiments 67-78, wherein the subject has received three or fewer prior lines of therapy. 80. Method according to one of embodiments 67-79, wherein the subject has received two or fewer prior lines of therapy. 81. Method according to one of embodiments 67-80, wherein the subject has received only one prior line of therapy. 82. Method according to one of embodiments 67-81, wherein the method is effective in obtaining an overall reaction rate (ORR) of about 75% to about 100%. 83. Method according to embodiment 82, wherein the ORR is approximately 84.6%. 84. Method according to one of embodiments 67-83, wherein the treatment is effective in maintaining the median progression-free survival (PFS) of the subject compared to administering DPd or PVd treatment. 85. Method according to embodiment 84, wherein the PFS after 12 months following administration of the treatment is approximately 75.9%. 86. Method according to embodiment 85, wherein the PFS at 12 months after administration of DPd or PVd is approximately 48.6%. 87. A method according to any embodiment 67-86, wherein the method further comprises treating the subject against a CAR-T-associated adverse event after administration of the dose of T cells, wherein the method optionally comprises administering treatment to the subject to alleviate the CAR-T-associated adverse event, wherein optionally the CAR-T-associated adverse event comprises cytokine release syndrome (CRS) and / or neurotoxicity, wherein optionally: (a) the CAR-T-associated adverse event is a CRS, wherein the CRS may further occur in the subject at a rate of approximately 60% to approximately 90% or at a rate of approximately 76.1%, wherein the maximum toxicity grade of the CRS may further be Grade 1, Grade 2 or Grade 3, wherein further: (1) the maximum toxicity level of the CRS is Grade 1, where, where applicable, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 52.8%; (2) the maximum toxicity level of the CRS is grade 2, where, where applicable, the maximum toxicity level of grade 2 occurs in the subject at a rate of approximately 22.2%; (3) the maximum toxicity level of the CRS is grade 3, where, where applicable, the maximum toxicity level of grade 3 occurs in the subject at a rate of approximately 1.1%; (4) the time to first onset of CRS is in the range of about 1 to about 23 days, with a median time to first onset of CRS of about 8 days where applicable; (5) the duration of the CRS ranges from about 1 to about 17 days, with a median duration of about 3 days where applicable; or (6) the treatment includes tocilizumab, oxygen, a corticosteroid, a vasopressor or any combination thereof; or (b) the CAR-T-associated adverse event is neurotoxicity, where appropriate the neurotoxicity includes an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, standard treatment-arm adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, where appropriate the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated neurotoxicity: (1) The immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of approximately 4.5%; (2) the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1 or Grade 2, wherein the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1, furthermore, where appropriate, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 3.4%, or where, where appropriate, the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 2, furthermore, where appropriate, the maximum toxicity level of Grade 2 occurs in the subject at a rate of approximately 1.1%; (3) the time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 6 to about 15 days, with a median time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom of about 9.5 days, if applicable; (4) the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 1 to about 6 days, with the median duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom being about 2 days; or (5) the treatment includes a corticosteroid and / or tocilizumab. 88. Method according to embodiment 87, wherein the neurotoxicity is CAR-T cell neurotoxicity, wherein optionally the CAR-T cell neurotoxicity occurs in the subject at a rate of about 17.0%, wherein optionally the CAR-T cell neurotoxicity comprises Grade 3 / 4 neurotoxicity, Grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, an adverse event occurring with exercise and neurocognitive treatment, or any combination thereof, further optionally comprising: (a) CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, occurring in approximately 2.3% of the subject; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, with cranial nerve palsy occurring in the subject at a rate of approximately 9.1%, where: (1) the cranial nerve palsy is grade 2 or grade 3, where furthermore, where appropriate, the cranial nerve palsy is grade 2, which occurs in the subject at a rate of about 8.0%, and where furthermore, where appropriate, the cranial nerve palsy is grade 3, which occurs in the subject at a rate of about 1.1%; (2) the time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject ranges from about 17 days to about 60 days, and furthermore, where applicable, the median time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject is about 21 days; (3) the cranial nerve palsy affects cranial nerve III, V or VII; (4) the duration of cranial nerve palsy ranges from about 15 days to about 262 days, with the median duration of cranial nerve palsy being about 77 days; or (5) the treatment includes a corticosteroid; (d) CAR T-cell neurotoxicity is a peripheral neuropathy, with peripheral neuropathy occurring in the subject at a rate of approximately 2.8%, if applicable; or (e) CAR-T cell neurotoxicity is an adverse event induced by exercise and neurocognitive treatment, wherein, where appropriate, the adverse event induced by exercise and neurocognitive treatment is grade 1, and furthermore, where, where appropriate, the grade 1 adverse event induced by neurocognitive treatment and the subject's adverse event induced by neurocognitive treatment occur at a rate of approximately 0.6%. 89. Method according to any embodiment 67-88, wherein the subject has one or more high-risk cytogenetic abnormalities selected from a group comprising the Gain / amp(1q), del(17p), t(4;14), t(14;16) or any combination thereof. 90. Method according to embodiment 89, wherein the subject has at least two cytogenetic abnormalities and wherein the subject optionally has two, three, four, five or more cytogenetic abnormalities. 91. Methods of embodiments 89 or 90, wherein the cytogenetic anomaly is a standard risk cytogenetic anomaly. 92. Method according to any embodiment 1-91, wherein the method further comprises treating the subject for a CAR-T-associated adverse event after administration of the dose of T cells, wherein the CAR-T-associated adverse event comprises cytokine release syndrome (CRS) and / or neurotoxicity. 93. Method according to embodiment 92, wherein the CRS CAR-T-associated adverse event is a CRS with a maximum toxicity level of grade 1, optionally wherein the maximum toxicity level of grade 1 occurs in the subject at a rate of about 52.8%. 94. Method according to embodiment 92, wherein the CRS-CAR-T-associated adverse event is a CRS with a maximum toxicity grade of 2, optionally wherein the maximum toxicity grade of 2 occurs in the subject at a rate of about 22.2%. 95. Method according to embodiment 92, wherein the CRS-CAR-T-associated adverse event is a CRS with a maximum toxicity grade of 3, optionally wherein the maximum toxicity grade of 3 occurs in the subject at a rate of about 1.1%. 96. Method according to one of embodiments 93-95, wherein the time until the first onset of the CRS is in the range of about 1 to about 23 days, optionally wherein the time until the first onset of the CRS is at a median of about 8 days. 97. Method according to one of embodiments 93-96, wherein the duration of the CRS is in the range of about 1 to about 17 days, optionally wherein the duration of the CRS is in a median of about 3 days. 98. Method according to any embodiment 93-97, wherein the treatment comprises tocilizumab, oxygen, a corticosteroid, a vasopressor or any combination thereof. 99. Method according to embodiment 92, wherein the CAR-T neurotoxicity-associated adverse event is selected from the group consisting of an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, treatment-related adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof. 100. Method according to embodiment 92, wherein the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated symptom. 101. Method according to embodiment 100, wherein the immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of about 4.5%. 102. Method according to embodiment 100 or 101, wherein the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom is grade 1, wherein the maximum toxicity grade 1 occurs in the subject at a rate of about 3.4%. 103. Method according to embodiment 100 or 101, wherein the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom is grade 2, wherein the maximum toxicity grade of grade 1 occurs in the subject at a rate of about 1.1%. 104. Method according to one of embodiments 100-103, wherein the time until the onset of the immune effector cell-associated neurotoxicity syndrome or associated symptom is in the range of about 6 to about 15 days. 105. Method according to embodiment 104, wherein the time to onset of the immune effector cell-associated neurotoxicity syndrome or associated symptom is on a median of about 9.5 days. 106. Method according to one of embodiments 100-103, wherein the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom is in the range of about 1 to about 6 days. 107. Method according to embodiment 106, wherein the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom is in a median of about 2 days. 108. Method according to one of embodiments 100-107, wherein the treatment comprises a corticosteroid and / or tocilizumab. 109. Method according to one of embodiments 99-108, wherein CAR-T cell neurotoxicity occurs in the subject at a rate of about 17.0%. 110. Method according to one of embodiments 99-109, wherein the CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity and wherein the grade 3 / 4 neurotoxicity occurs in the subject at a rate of about 2.3%. 111. Method according to one of embodiments 99-109, wherein the CAR-T cell neurotoxicity is grade 5 neurotoxicity. 112. Method according to one of embodiments 99-109, wherein the CAR-T cell neurotoxicity is cranial nerve palsy and wherein the cranial nerve palsy occurs in the subject at a rate of about 9.1%. 113. Method according to embodiment 112, wherein the cranial nerve palsy is grade 2 cranial nerve palsy, wherein grade 2 cranial nerve palsy occurs in the subject at a rate of about 8.0%. 114. Method according to embodiment 112, wherein the cranial nerve palsy is grade 3 cranial nerve palsy, wherein grade 2 cranial nerve palsy occurs in the subject at a rate of about 1.1%. 115. Method according to one of embodiments 112-114, wherein the time until the onset of cranial nerve paralysis after administration of the dose of T cells to the subject is in the range of about 17 days to about 60 days. 116. Method according to one of embodiments 112-115, wherein the cranial nerve palsy affects cranial nerve III, V or VII 117. Method according to one of embodiments 112-116, wherein the duration of cranial nerve paralysis is in the range of about 15 days to about 262 days. 118. Method according to one of embodiments 112-117, wherein the treatment comprises a corticosteroid. 119. Method according to one of embodiments 99-109, wherein the CAR-T cell neurotoxicity is peripheral neuropathy and wherein the peripheral neuropathy occurs in the subject at a rate of about 2.8%. 120. Method according to one of embodiments 99-109, wherein CAR-T cell neurotoxicity is an adverse event (MNT) induced by movement and neurocognitive treatment. 121. Method according to embodiment 120, wherein the MNT is an MNT grade 1, wherein the MNT grade 1 occurs in the subject at a rate of about 0.6%. 122. Method according to embodiment 64 or 89, wherein the cytogenetic anomaly comprises Gain / amp(1q). 123. Method according to embodiment 64 or 89, wherein the cytogenetic anomaly comprises del(17p). 124. Method according to embodiment 64 or 89, wherein the cytogenetic anomaly comprises t(4;14). 125. Method according to embodiment 64 or 89, wherein the cytogenetic anomaly comprises t(14;16). 126. Method according to one of embodiments 1-91, wherein the subject has a reduced risk of developing a CAR-T-associated adverse event. 127. Method according to embodiment 126, wherein the CAR-T-associated adverse event comprises a cytokine release syndrome (CRS). 128. Method according to embodiment 127, wherein the CRS occurs in the subject at a rate of about 60% to about 90%, or at a rate of about 76.1%. 129. Method according to embodiment 127 or 128, wherein the maximum toxicity grade of the CRS is Grade 1, Grade 2 or Grade 3. 130. Method according to any embodiment 126 to 129, wherein the maximum toxicity level of the CRS is Grade 1, wherein, optionally, the maximum toxicity level of Grade 1 occurs in the subject at a rate of about 52.8%. 131. Method according to any embodiment 126 to 130, wherein the maximum toxicity level of the CRS is grade 2, optionally wherein the maximum toxicity level of grade 2 occurs in the subject at a rate of about 22.2%. 132. Method according to any embodiment 126 to 131, wherein the maximum toxicity level of the CRS is grade 3, wherein, optionally, the maximum toxicity level of grade 3 occurs in the subject at a rate of about 1.1%. 133. Method according to embodiment 126, wherein the CAR-T-associated adverse event comprises neurotoxicity. 134. Method according to embodiment 133, wherein the neurotoxicity CAR-T-associated adverse event is selected from the group consisting of an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, treatment-related adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof. 135. Method according to embodiment 134, wherein the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated symptom. 136. Method according to embodiment 135, wherein the immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of about 4.5%. 137. Method according to embodiment 135 or 136, wherein the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom is grade 1, wherein the maximum toxicity grade 1 occurs in the subject at a rate of about 3.4%. 138. Method according to embodiment 135 or 136, wherein the maximum toxicity grade of the immune effector cell-associated neurotoxicity syndrome or associated symptom is grade 2, wherein the maximum toxicity grade of grade 1 occurs in the subject at a rate of about 1.1%. 139. Method according to one of embodiments 135-138, wherein the time until the onset of the immune effector cell-associated neurotoxicity syndrome or associated symptom is in the range of about 6 to about 15 days. 140. Method according to embodiment 139, wherein the time to onset of the immune effector cell-associated neurotoxicity syndrome or associated symptom is on a median of about 9.5 days. 141. Method according to one of embodiments 135-138, wherein the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom is in the range of about 1 to about 6 days. 142. Method according to embodiment 141, wherein the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom is in a median of about 2 days. 143. Method according to one of embodiments 135-142, wherein the treatment comprises a corticosteroid and / or tocilizumab. 144. Method according to one of embodiments 134-143, wherein CAR-T cell neurotoxicity occurs in the subject at a rate of about 17.0%. 145. Method according to one of embodiments 134-144, wherein the CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity and wherein the grade 3 / 4 neurotoxicity occurs in the subject at a rate of about 2.3%. 146. Method according to one of embodiments 134-145, wherein the CAR-T cell neurotoxicity is grade 5 neurotoxicity. 147. Method according to one of embodiments 134-144, wherein the CAR-T cell neurotoxicity is cranial nerve palsy and wherein the cranial nerve palsy occurs in the subject at a rate of about 9.1%. 148. Method according to embodiment 147, wherein the cranial nerve palsy is grade 2 cranial nerve palsy, wherein grade 2 cranial nerve palsy occurs in the subject at a rate of about 8.0%. 149. Method according to embodiment 147, wherein the cranial nerve palsy is grade 3 cranial nerve palsy, wherein grade 2 cranial nerve palsy occurs in the subject at a rate of about 1.1%. 150. Method according to one of embodiments 147-149, wherein the time until the onset of cranial nerve paralysis after administration of the dose of T cells to the subject is in the range of about 17 days to about 60 days. 151. Method according to one of embodiments 147-150, wherein the cranial nerve palsy affects cranial nerve III, V or VII 152. Method according to one of embodiments 147-151, wherein the duration of cranial nerve paralysis is in the range of about 15 days to about 262 days. 153. Method according to one of embodiments 147-152, wherein the treatment comprises a corticosteroid. 154. Method according to one of embodiments 134-144, wherein the CAR-T cell neurotoxicity is peripheral neuropathy and wherein the peripheral neuropathy occurs in the subject at a rate of about 2.8%. 155. Method according to one of embodiments 134-144, wherein CAR-T cell neurotoxicity is an adverse event (MNT) induced by movement and neurocognitive treatment. 156. Method according to embodiment 155, wherein the MNT is an MNT grade 1, wherein the MNT grade 1 occurs in the subject at a rate of about 0.6%. 157. Method according to one of embodiments 38, 63, 88, 119 and 154, wherein the peripheral neuropathy is grade 1 and wherein the peripheral neuropathy grade 1 occurs at a rate of about 1.1%. 158. Method according to one of embodiments 38, 63, 88, 119 and 154, wherein the peripheral neuropathy is grade 2 and wherein the peripheral neuropathy grade 2 occurs at a rate of about 1.1%. 159. Method according to one of embodiments 38, 63, 88, 119 and 154, wherein the peripheral neuropathy is grade 3 and wherein the peripheral neuropathy grade 3 occurs at a rate of about 0.6%. 160. Method for the selective treatment of a subject, comprising administering a dose of T cells comprising a chimeric antigen receptor (CAR) to the subject in whom a high-risk feature has been identified, comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of BCMA, (b) a transmembrane domain, and (c) an intracellular signaling domain, where the CAR comprises the amino acid sequence of SEQ ID NO: 17, where the high-risk feature is a cytogenetic abnormality, an International Staging System (ISS) stage III and / or soft tissue plasmacytomas, where the subject has multiple myeloma, has received one to three prior lines of therapy, including therapy with an IMiD, and is refractory to the IMiD,

[0244] Whereby the method is effective in obtaining an overall reaction as described in Example 3, one of Tables 1-5 and 15, or one of the Fig. 6A-6C, 7 and 8A-8D, The procedure may further include treating the subject for an adverse event following administration of the dose of T cells as described in Example 4 or one of Tables 6-14 and 16, and The procedure may further include administering treatment to the subject to alleviate the undesired event, as described in Example 4, or in one of Tables 6-14 and 16. EXAMPLES

[0245] The following examples are provided to further describe some of the aspects and embodiments disclosed herein. The examples are intended to illustrate, not limit, the disclosed aspects or embodiments. Example 1: Ciltacabtagene Autoleucel

[0246] B-cell maturation antigen (BCMA, also known as CD269 and TNFRSF17) is a 20-kilodalton, type III membrane protein belonging to the tumor necrosis receptor superfamily. BCMA is a cell surface antigen that is predominantly expressed at high levels in B-cell lineage cells. Fig.Figure 1 shows the expression of BCMA on various immune-derived cells. Comparative studies have shown that BCMA is not present in most normal tissues and is not expressed on CD34-positive hematopoietic stem cells. BCMA binds to ligands that induce B-cell proliferation and plays a critical role in B-cell maturation and subsequent differentiation into plasma cells. Its selective expression and biological significance for myeloma cell proliferation and survival make BCMA a promising target for CAR-T cell-based immunotherapy.

[0247] Ciltacabtagene autoleucel (Cilta-cel) is an autologous chimeric antigen receptor T-cell (CAR-T) therapy targeting BCMA. The chimeric ciltacabtagene autoleucel antigen receptor (CAR) comprises two B-cell maturation antigen (BCMA)-targeting VHH domains designed for avidity. A map of the construct is shown in Fig.Figure 2 shows a scheme of CAR-T cell production. Fig. Figure 3 shows that Cilta-Cell includes a VHH domain comprising the amino acid sequence of SEQ ID NO: 2 and a VHH domain comprising the amino acid sequence of SEQ ID NO: 4. Example 2: Treatment procedure with ciltacabtagene autoleucel

[0248] Cilta-Cell is highly effective in heavily pretreated relapsed / refractory multiple myeloma (RRMM). In this study, we investigated Cilta-Cell as an early line of treatment in lenalidomide-refractory patients.

[0249] Most patients with multiple myeloma (MM) relapse after standard treatment (van de Donk, Hematology Am Soc Hematol Educ Program 2020;2020:248-58; Rodriguez-Lobato et al., Br J Haematol 2022;196:649-59) and outcomes worsen with each subsequent line of therapy (LOT) (Yong et al., Br J Haematol 2016;175:252-64; Dhakal et al., Clinical Lymphoma Myeloma and Leukemia 2022;22:S167; Dhakal et al., HemaSphere 2022;6:790-1). Lenalidomide is an immunomodulator recommended for newly diagnosed and relapsed / refractory multiple myeloma (RRMM) (Dimopoulos et al., HemaSphere 2021;5:e528; National Comprehensive Cancer Network. NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines®) Version 3, 2023). The use of lenalidomide is widespread in early settings, including as maintenance therapy (van de Donk, Hematology am Soc Hematol Educ Program 2020;2020:248-58; de Arriba de la Fuente et al., Cancer (Basel) 2022;15).Lenalidomide refractoriness rates at the start of patients' treatment journeys are increasing (van de Donk, Hematology Am Soc Hematol Educ Program 2020;2020:248-58; de Arriba de la Fuente et al., Cancer (Basel) 2022;15), leading to a growing need for new, effective therapies for lenalidomide-refractory disease (de Arriba de la Fuente et al.; Cancer (Basel) 2022;15). A high treatment discontinuation rate—only 13% to 35% of patients receive ≥4 lot sizes—also highlights the need to initiate effective therapies early (Fonseca et al., BMC Cancer 2020;20:1087).

[0250] Cilta-cel led to early, deep, and durable responses in patients with RRMM and ≥3 prior CHARGE in the phase 1b / 2 CARTITUDE-1 trial (median progression-free survival [PFS], 34.9 months) (Berdeja et al., Lancet 2021;398:314-24; Martin et al., J Clin Oncol 2022:JCO2200842; Lin et al., J Clin Oncol 2023. In submission). The Phase 2 CARTITUDE-2 study (cohorts A and B) demonstrated the efficacy of Cilta-cel in small cohorts in earlier stages of disease with response rates of 95%-100%, and median duration of response (DOR) and median progression-free survival (PFS) in 2022, which were not reached after approximately 1.5 years of follow-up (van de Donk et al., Blood 2022;140:7536-7; Einsele et al., American Society of Clinical Meeting; IL, June 3-7;).

[0251] CARTITUDE-4 is a randomized, controlled phase 3 trial comparing Cilta-cel with physician choice between two highly effective standard therapies in patients with lenalidomide-refractory multiple myeloma after 1-3 batches. We report efficacy and safety results from the first planned analysis of CARTITUDE-4. Study design and patients

[0252] CARTITUDE-4 is a global, open-label, randomized, phase 3 trial conducted at 81 sites in the US, Europe, Asia, and Australia. Eligible patients were lenalidomide-refractory (Rajkumar et al., Blood 2011;117:4691-5), had received 1-3 prior CHARGE therapies, including a proteasome inhibitor and an immunomodulatory drug, had an Eastern Cooperative Oncology Group performance status score of ≤1, no prior CAR-T cell therapy, and no prior BCMA-targeted treatment. Randomization and treatments

[0253] Patients were randomized 1:1 by computer-generated randomization to standard therapy (physician-selected combination of pomalidomide-bortezomib-dexamethasone [PVd] (Richardson et al., Lancet Oncol 2019; 20:781-94) or daratumumab-pomalidomide-dexamethasone [DPd]) (Dimopoulos et al., Lancet Oncol 2021;22:801-12) or a single Cilta-cel infusion following physician-selected bridging therapy (PVd or DPd). Randomization was stratified by choice of PVd versus DPd, International Staging System (ISS) stage at screening (I vs. II vs. III), and number of previous batches (1 vs. 2-3).

[0254] In the standard treatment arm, DPd was administered in 28-day cycles and PVd in 21-day cycles until disease progression. Patients in the Cilta-cel arm underwent apheresis, followed by ≥1 bridging therapy cycle (number of cycles based on clinical status and Cilta-cel manufacturing time) and lymphodepleti...

Claims

[1] Dose of T cells for use in a procedure for the treatment of multiple myeloma in a subject, wherein the T cells comprise a chimeric antigen receptor (CAR), comprising: (a) an extracellular antigen-binding domain capable of binding specifically to an epitope of the B-cell maturation antigen (BCMA), (b) a transmembrane domain, and (c) an intracellular signaling domain, wherein the subject has received one to three prior lines of therapy, including therapy with an immunomodulatory drug (IMiD), and is refractory to the IMiD. [2] The dose of T cells for use according to claim 1, wherein the subject has a high-risk feature, and wherein the high-risk feature may optionally be a cytogenetic abnormality, an International Staging System (ISS) stage III and / or soft tissue plasmacytomas. [3] Dose of T cells for use in a procedure for selectively treating a subject, wherein the T cells comprise a chimeric antigen receptor (CAR), comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of BCMA, (b) a transmembrane domain, and (c) an intracellular signaling domain, the procedure includes: (1) Determine whether the subject has a high-risk feature, wherein the high-risk feature is a cytogenetic abnormality, an International Staging System (ISS) stage III and / or soft tissue plasmacytomas; and (2) Administering the dose of T cells to the subject determined to have the high-risk characteristic in step (1). where the subject may have multiple myeloma, has received one to three prior lines of therapy, including therapy with an IMiD, and is refractory to the IMiD. [4] Dose of T cells for use in a procedure for the selective treatment of a subject, wherein the T cells comprise a chimeric antigen receptor (CAR), comprising: (a) an extracellular antigen-binding domain capable of specifically binding to an epitope of BCMA, (b) a transmembrane domain, and (c) an intracellular signaling domain, the procedure involves administering the dose of T cells to the subject in whom a high-risk characteristic has been identified, where the high-risk feature is a cytogenetic abnormality, an International Staging System (ISS) stage III and / or soft tissue plasmacytomas, where the subject may have multiple myeloma, has received one to three prior lines of therapy, including therapy with an IMiD, and is refractory to the IMiD. [5] The dose of T cells for use according to any one of claims 1-4, wherein the IMiD is lenalidomide. [6] The dose of T cells for use according to any one of claims 2-5, wherein the high-risk feature is a cytogenetic abnormality. [7] The dose of T cells for use according to claim 6, wherein the cytogenetic abnormality is a high-risk cytogenetic abnormality. [8] The dose of T cells for use according to claim 7, wherein the subject has one or more high-risk cytogenetic abnormalities selected from a group comprising Gain / AMP(1q), del(17p), t(4;14), t(14;16) or any combination thereof. [9] The dose of T cells for use according to claim 8, wherein the cytogenetic anomaly comprises Gain / amp(1q). [10] The dose of T cells for use according to claim 8, wherein the cytogenetic abnormality comprises del(17p). [11] The dose of T cells for use according to claim 8, wherein the cytogenetic abnormality comprises t(4;14). [12] The dose of T cells for use according to claim 8, wherein the cytogenetic abnormality comprises t(14;16). [13] The dose of T cells for use according to any one of claims 6-12, wherein the subject has at least two cytogenetic abnormalities and wherein the subject optionally has two, three, four, five or more cytogenetic abnormalities. [14] The dose of T cells for use according to claim 6, wherein the cytogenetic abnormality is a standard risk cytogenetic abnormality. [15] The dose of T cells for use according to any one of claims 2-5, wherein the high-risk feature is International Staging System (ISS) level III. [16] The dose of T cells for use according to any one of claims 2-5, wherein the high-risk feature is soft tissue plasmacytomas. [17] The dose of T cells for use according to any one of claims 1-16, wherein the subject has received a prior line of therapy. [18] The dose of T cells for use according to any one of claims 1-16, wherein the subject has received two prior lines of therapy. [19] The dose of T cells for use according to any one of claims 1-16, wherein the subject has received three prior lines of therapy. [20] The dose of T cells for use according to any one of claims 1-19, wherein the one, two or three prior lines of therapy include treatment with pomalidomide. [21] The dose of T cells for use according to any one of claims 1-20, wherein the one, two or three prior lines of therapy further comprise treatment with an anti-CD38 antibody and wherein the anti-CD38 antibody is optionally daratumumab and / or isatuximab. [22] The dose of T cells for use according to any one of claims 1-21, wherein the one, two or three prior lines of therapy further comprise treatment with a proteasome inhibitor, wherein the proteasome inhibitor is optionally bortezomib, carfilzomib, ixazomib or any combination thereof. [23] The dose of T cells for use according to any one of claims 1-22, wherein the subject has further received bridge therapy, wherein the bridge therapy is optionally the choice of the physician, wherein the bridge therapy optionally comprises pomalidomide, bortezomib, dexamethasone, daratumumab or any combination thereof, wherein the bridge therapy optionally comprises pomalidomide, bortezomib and dexamethasone, and wherein the bridge therapy optionally comprises daratumumab, pomalidomide and dexamethasone. [24] The dose of T cells for use according to claim 23, wherein the subject has received the bridge therapy from about every 20 days to about every 30 days, wherein the subject may optionally receive the bridge therapy about every 21 days, wherein the subject may optionally receive the bridge therapy about every 28 days and wherein the subject may further optionally receive at least one, two, three, four or more bridge therapies. [25] The dose of T cells for use according to any one of claims 1 to 24, wherein the subject has additionally received lymphodepletion therapy, wherein the lymphodepletion therapy optionally comprises cyclophosphamide and / or fludarabine daily, wherein the lymphodepletion therapy optionally comprises cyclophosphamide and fludarabine daily, wherein the lymphodepletion treatment optionally comprises cyclophosphamide at a concentration of about 300 mg / m² 2 and fludarabine at a concentration of approximately 30 mg / m³ 2 daily over 3 days. [26] The dose of T cells for use according to any one of claims 1-25, wherein the dose of T cells is 0.5-1.0 × 10 6 -cells / kg body weight of the subject, where, if necessary, the dose of T cells is approximately 0.75 × 10 6-cells / kg body weight of the subject, the procedure optionally comprising administering the dose of T cells approximately 5 to approximately 7 days after the start of lymphodepletion therapy, optionally administering the dose as a single infusion. [27] The dose of T cells for use according to any one of claims 1-26, wherein the method is effective in maintaining an overall response in the subject after administration of the dose of T cells to the subject, wherein optionally the method is effective in maintaining the overall response at a rate of about 75% to about 100%, wherein further optionally the method is effective in maintaining the overall response at a rate of about 84.6%, and wherein further optionally the method is effective in maintaining the overall response at a rate of about 99.4%. [28] The dose of T cells for use according to claim 27, wherein the overall response, in order from best to worst, comprises: (1) a stringent and complete response; (2) a complete reaction; (3) a very good partial response; (4) a partial reaction; or (5) a minimal response. [29] The dose of T cells for use according to claim 28, wherein the overall response is a stringent complete response, wherein the method is optionally effective in obtaining the stringent complete response at a rate of about 40% to about 90%, about 50% to about 80%, about 58.2% or about 68.8%. [30] The dose of T cells for use according to claim 28, wherein the overall reaction is a complete reaction, wherein optionally the method is effective to obtain the complete reaction at a rate of about 10% to about 20%, and further optionally the method is effective to obtain the complete reaction at a rate of about 14.9% or about 17.6%. [31] The dose of T cells for use according to claim 28, wherein the overall response is a very good partial response or a partial response. [32] The dose of T cells for use according to claim 28, wherein: (1) the process is effective in obtaining a stringent complete reaction or a complete reaction at a rate of about 70% to about 90%, and optionally the process is effective in obtaining a stringent complete reaction or a complete reaction at a rate of about 73.1% or about 86.4%; (2) the process is effective in obtaining a stringent complete reaction, a complete reaction or a very good partial reaction at a rate of about 80% to about 100%, wherein the process is optionally effective in obtaining a stringent complete reaction, a complete reaction or a very good partial reaction at a rate of about 81.3% or about 96.0%; (3) the procedure is effective in obtaining a minimal response; (4) the procedure is effective for maintaining a minimal residual disease negative, where the procedure may be effective for maintaining a minimal residual disease negative at a rate of approximately 50% to approximately 80%, where the procedure may continue to be effective for maintaining a minimal residual disease negative at a rate of approximately 60.6% or approximately 71.6%; or (5) the procedure is effective in maintaining progression-free 12-month survival for at least about 60% to about 100% of subjects, at least about 69.4% to about 81.1% of subjects or at least about 84.1% to about 93.4% of subjects, where appropriate the procedure is effective in maintaining progression-free 12-month survival for at least about 75.9% of subjects or about 89.7% of subjects. [33] The dose of T cells for use according to any one of claims 27-32, wherein: (1) the time to the first overall response or first minimal response is in the range of approximately 0.9 to approximately 11.1 months, with the median time to the first overall response or first minimal response being approximately 2.1 months; or (2) the time to best overall response or best minimum response is approximately 1.1 to approximately 18.6 months, with the median time to best overall response or best minimum response being approximately 6.4 or approximately 6.5 months, where appropriate. [34] The dose of T cells for use according to any one of claims 1-33, wherein the method further comprises treating the subject for an adverse event following administration of the dose of T cells, wherein the method optionally comprises administering treatment to the subject to alleviate the adverse event, wherein optionally the adverse event comprises a hematological adverse event, a non-hematological adverse event, a treatment-related adverse event, or any combination thereof, wherein optionally the non-hematological adverse event comprises an infection other than an infection and / or a non-hematological adverse event other than an infection, wherein optionally the adverse event comprises neutropenia, thrombocytopenia, anemia, lymphopenia, an upper respiratory tract infection, nasopharyngitis, sinusitis, rhinitis, tonsillitis, pharyngitis, laryngitis, pharyngotonsillitis,COVID-19, COVID-19 pneumonia, asymptomatic COVID-19, neutropenic sepsis, progressive multifocal leukoencephalopathy, septic shock, respiratory failure, pulmonary embolism, lower respiratory tract / lung infection, pneumonia, bronchitis, nausea, hypogammaglobulinemia, diarrhea, fatigue, headache, constipation, hypokalemia, asthenia, peripheral edema, decreased appetite, peripheral sensory neuropathy, back pain, arthralgia, pyrexia, dyspnea, insomnia, or any combination thereof, where the adverse event is, where appropriate, a Grade 3 / 4 adverse event and where, where appropriate, the adverse event lasts for more than approximately 30 days or approximately 60 days. [35] The dose of T cells for use according to any one of claims 1-34, wherein the method further comprises treating the subject for a second primary malignancy after administering the dose of T cells, wherein the method optionally comprises administering a treatment to the subject to alleviate the second primary malignancy, wherein optionally the second primary malignancy comprises a cutaneous / non-invasive malignancy, a hematological malignancy, a non-cutaneous / invasive malignancy, or any combination thereof, wherein optionally the second primary malignancy comprises basal cell carcinoma, Bowen's disease, lip squamous cell carcinoma, malignant melanoma, malignant melanoma in situ, cutaneous squamous cell carcinoma, acute myeloid leukemia, a myelodysplastic syndrome, peripheral T-cell lymphoma, angiosarcoma, invasive lobular breast carcinoma, pleomorphic malignant fibrous Histiocytoma, renal cell carcinoma,Tonsil carcinoma or any combination thereof. [36] The dose of T cells for use according to claim 34 or 35, wherein the adverse event or second primary malignancy occurs in the subject at a rate comparable to the rate of the same adverse event or second primary malignancy occurring in a subject undergoing standard treatment. [37] The dose of T cells for use according to any one of claims 1-36, wherein the method further comprises treating the subject against a CAR-T-associated adverse event after administration of the dose of T cells, wherein the method optionally comprises administering a treatment to the subject to alleviate the CAR-T-associated adverse event, wherein optionally the CAR-T-associated adverse event comprises cytokine release syndrome (CRS) and / or neurotoxicity, wherein optionally: (a) the CAR-T-associated adverse event is a CRS, wherein the CRS may further occur in the subject at a rate of approximately 60% to approximately 90% or at a rate of approximately 76.1%, wherein the maximum toxicity grade of the CRS may further be Grade 1, Grade 2 or Grade 3, wherein further: (1) the maximum toxicity level of the CRS is Grade 1, where, where applicable, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 52.8%; (2) the maximum toxicity level of the CRS is grade 2, where, where applicable, the maximum toxicity level of grade 2 occurs in the subject at a rate of approximately 22.2%; (3) the maximum toxicity level of the CRS is grade 3, where, where applicable, the maximum toxicity level of grade 3 occurs in the subject at a rate of approximately 1.1%; (4) the time to first onset of CRS is in the range of about 1 to about 23 days, with a median time to first onset of CRS of about 8 days where applicable; (5) the duration of the CRS ranges from about 1 to about 17 days, with a median duration of about 3 days where applicable; or (6) the treatment includes tocilizumab, oxygen, a corticosteroid, a vasopressor or any combination thereof; or (b) the CAR-T-associated adverse event is neurotoxicity, where appropriate the neurotoxicity includes an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, standard treatment-arm adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, where appropriate the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated neurotoxicity: (1) The immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of approximately 4.5%; (2) the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1 or Grade 2, wherein the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1, furthermore, where appropriate, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 3.4%, or where, where appropriate, the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 2, furthermore, where appropriate, the maximum toxicity level of Grade 2 occurs in the subject at a rate of approximately 1.1%; (3) the time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 6 to about 15 days, with a median time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom of about 9.5 days, if applicable; (4) the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 1 to about 6 days, with the median duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom being about 2 days; or (5) the treatment includes a corticosteroid and / or tocilizumab. [38] The dose of T cells for use according to claim 37, wherein the neurotoxicity is CAR-T cell neurotoxicity, wherein optionally the CAR-T cell neurotoxicity occurs in the subject at a rate of about 17.0%, wherein optionally the CAR-T cell neurotoxicity comprises grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, a movement event and a neurocognitive treatment-related adverse event or any combination thereof, wherein further optionally: (a) CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, occurring in approximately 2.3% of the subject; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, with cranial nerve palsy occurring in the subject at a rate of approximately 9.1%, where: (1) the cranial nerve palsy is grade 2 or grade 3, where furthermore, where appropriate, the cranial nerve palsy is grade 2, which occurs in the subject at a rate of about 8.0%, and where furthermore, where appropriate, the cranial nerve palsy is grade 3, which occurs in the subject at a rate of about 1.1%; (2) the time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject ranges from about 17 days to about 60 days, and furthermore, where applicable, the median time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject is about 21 days; (3) the cranial nerve palsy affects cranial nerve III, V or VII; (4) the duration of cranial nerve palsy ranges from about 15 days to about 262 days, with the median duration of cranial nerve palsy being about 77 days; or (5) the treatment includes a corticosteroid; (d) CAR T-cell neurotoxicity is a peripheral neuropathy, with peripheral neuropathy occurring in the subject at a rate of approximately 2.8%, if applicable; or (e) CAR-T cell neurotoxicity is an adverse event induced by exercise and neurocognitive treatment, wherein, where appropriate, the adverse event induced by exercise and neurocognitive treatment is grade 1, and furthermore, where, where appropriate, the grade 1 adverse event induced by neurocognitive treatment and the subject's adverse event induced by neurocognitive treatment occur at a rate of approximately 0.6%. [39] The dose of T cells for use according to any one of claims 1-38, wherein: (a) CD3+ cells encompassing the CAR in the subject's blood reach their median peak approximately 13 days after administration of the T cells to the subject, with the CD3+ cells encompassing the CAR in the subject's blood potentially reaching a peak at a mean concentration of approximately 1523 cells / µl; (b) CD3+ cells encompassing the CAR remain detectable in the subject's blood from approximately 13 days to approximately 631 days after administration of the T cells to the subject, with the CD3+ cells encompassing the CAR remaining detectable in the subject's blood for a median of approximately 57 days after administration of the T cells to the subject, if applicable; or (c) the AUC 0-28 The average number of CD3+ cells encompassing the CAR in the subject's blood is approximately 12,504 cells / µL. [40] The dose of T cells for use according to any one of claims 1-39, wherein the first VHH domain comprises a CDR1, a CDR2 and a CDR3 as shown in the VHH domain, comprising the amino acid sequence of SEQ ID NO: 2, and the second VHH domain comprises a CDR1, a CDR2 and a CDR3 as shown in the VHH domain, comprising the amino acid sequence of SEQ ID NO: 4, wherein optionally the first VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 18, a CDR2 comprising the amino acid sequence of SEQ ID NO: 19, a CDR3 comprising the amino acid sequence of SEQ ID NO: 20, and the second VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a CDR2 comprising the amino acid sequence of SEQ ID NO: 22, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 23.wherein optionally the first VHH domain comprises the amino acid sequence of SEQ ID NO: 2 and the second VHH domain comprises the amino acid sequence of SEQ ID NO: 4, wherein optionally the first VHH domain is located at the N-terminus of the second VHH domain or the first VHH domain is located at the C-terminus of the second VHH domain, wherein further optionally:, (a) the first VHH domain is linked to the second VHH domain via a linker comprising the amino acid sequence of SEQ ID NO: 3; (b) the transmembrane domain is derived from a molecule selected from the group consisting of CD8α, CD4, CD28, CD137, CD80, CD86, CD152 and PD1, optionally the transmembrane domain being derived from CD8α and the amino acid sequence being SEQ ID NO: 6; (c) the intracellular signaling domain comprises a primary intracellular signaling domain of an immune effector cell, the primary intracellular signaling domain optionally being derived from CD3ζ, comprising the amino acid sequence of SEQ ID NO: 8; (d) the intracellular signaling domain comprises a costimulatory signaling domain, wherein optionally the costimulatory signaling domain is derived from a costimulatory molecule selected from the group consisting of CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, ligands of CD83 and any combination thereof, wherein optionally the costimulatory signaling domain comprises a cytoplasmic domain of CD137 comprising the amino acid sequence of SEQ ID NO: 7; (e) the CAR further comprises a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain, the hinge domain optionally being derived from CD8α, comprising the amino acid sequence of SEQ ID NO: 5; (f) the CAR further comprises a signal peptide located at the N-terminus of the polypeptide, the signal peptide optionally being derived from CD8α, comprising the amino acid sequence of SEQ ID NO: 1; or (g) of the CAR comprises the amino acid sequence of SEQ ID NO:

17. [41] The dose of T cells for use according to any one of claims 1-40, wherein the dose of T cells is formulated in a composition comprising 5% dimethyl sulfoxide (DMSO). [42] The dose of T cells for use according to any one of claims 1-41, wherein the administration of the dose of T cells reduces the risk of disease progression or death in the subject. [43] The dose of T cells for use according to claim 42, wherein the risk of disease progression or death is reduced compared to treatment with daratumumab pomalidomide dexamethasone (DPd) or pomalidomide bortezomib dexamethasone (PVd). [44] The dose of T cells for use according to claim 42, wherein the risk of disease progression or death is reduced compared to administering a double-cell treatment. [45] The dose of T cells for use according to one of claims 42-44, wherein the subject has an approximately 60% to approximately 75% reduced risk of disease progression or death. [46] The dose of T cells for use according to claim 45, wherein the subject has an approximately 74% reduced risk of disease progression or death. [47] The dose of T cells for use according to any one of claims 42-46, wherein the IMiD is lenalidomide. [48] ​​The dose of T cells for use according to any one of claims 42-47, wherein the subject has received three or fewer prior lines of therapy. [49] The dose of T cells for use according to any one of claims 42-47, wherein the subject has received two or fewer prior lines of therapy. [50] The dose of T cells for use according to any one of claims 42-47, wherein the subject has received only one prior line of therapy. [51] The dose of T cells for use according to any one of claims 42-50, wherein the method is effective in obtaining an overall response rate (ORR) of about 75% to about 100%. [52] The dose of T cells for use according to claim 51, wherein the ORR is about 84.6%. [53] The dose of T cells for use according to any one of claims 42-52, wherein the treatment is effective in maintaining the median progression-free survival (PFS) of the subject compared to administration of DPd or PVd treatment. [54] The dose of T cells for use according to claim 53, wherein the PFS after 12 months following administration of the treatment is about 75.9%. [55] The dose of T cells for use according to claim 53 or 54, wherein the PFS 12 months after administration of DPd or PVd is about 48.6%. [56] The dose of T cells for use according to one of claims 1-23 and 42-55, wherein the treatment is more effective in obtaining a stringent complete response (sCR) in the subject compared to the administration of a DPd or PVd treatment. [57] The dose of T cells for use according to claim 56, wherein the sCR after administration of the treatment is about 58.2%. [58] The dose of T cells for use according to claim 56 or 57, wherein the sCR after administration of DPd or PVd is about 15.2%. [59] The dose of T cells for use according to one of claims 1-23 and 42-58, wherein the treatment is more effective in achieving a very good partial response (VGPR) or better in the subject compared to the administration of a DPd or PVd treatment. [60] The dose of T cells for use according to claim 59, wherein the VGPR or better after administration of the treatment is about 81.3%. [61] The dose of T cells for use according to claim 59 or 60, wherein the VGPR or better after administration of DPd or PVd is about 45.5%. [62] The dose of T cells for use according to any one of claims 42-61, wherein the method further comprises treating the subject against a CAR-T-associated adverse event after administration of the dose of T cells, wherein the method optionally comprises administering a treatment to the subject to alleviate the CAR-T-associated adverse event, wherein optionally the CAR-T-associated adverse event comprises cytokine release syndrome (CRS) and / or neurotoxicity, wherein optionally: (a) the CAR-T-associated adverse event is a CRS, wherein the CRS may further occur in the subject at a rate of approximately 60% to approximately 90% or at a rate of approximately 76.1%, wherein the maximum toxicity grade of the CRS may further be Grade 1, Grade 2 or Grade 3, wherein further: (1) the maximum toxicity level of the CRS is Grade 1, where, where applicable, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 52.8%; (2) the maximum toxicity level of the CRS is grade 2, where, where applicable, the maximum toxicity level of grade 2 occurs in the subject at a rate of approximately 22.2%; (3) the maximum toxicity level of the CRS is grade 3, where, where applicable, the maximum toxicity level of grade 3 occurs in the subject at a rate of approximately 1.1%; (4) the time to first onset of CRS is in the range of about 1 to about 23 days, with a median time to first onset of CRS of about 8 days where applicable; (5) the duration of the CRS ranges from about 1 to about 17 days, with a median duration of about 3 days where applicable; or (6) the treatment includes tocilizumab, oxygen, a corticosteroid, a vasopressor or any combination thereof; or (b) the CAR-T-associated adverse event is neurotoxicity, where appropriate the neurotoxicity includes an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, standard treatment-arm adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, where appropriate the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated neurotoxicity: (1) The immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of approximately 4.5%; (2) the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1 or Grade 2, wherein the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1, furthermore, where appropriate, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 3.4%, or where, where appropriate, the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 2, furthermore, where appropriate, the maximum toxicity level of Grade 2 occurs in the subject at a rate of approximately 1.1%; (3) the time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 6 to about 15 days, with a median time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom of about 9.5 days, if applicable; (4) the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 1 to about 6 days, with the median duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom being about 2 days; or (5) the treatment includes a corticosteroid and / or tocilizumab. [63] The dose of T cells for use according to claim 62, wherein the neurotoxicity is CAR-T cell neurotoxicity, wherein the CAR-T cell neurotoxicity optionally occurs in the subject at a rate of about 17.0%, wherein the CAR-T cell neurotoxicity optionally comprises grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, a movement event and a neurocognitive treatment-related adverse event or any combination thereof, further optionally comprising: (a) CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, occurring in approximately 2.3% of the subject; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, with cranial nerve palsy occurring in the subject at a rate of approximately 9.1%, where: (1) the cranial nerve palsy is grade 2 or grade 3, where furthermore, where appropriate, the cranial nerve palsy is grade 2, which occurs in the subject at a rate of about 8.0%, and where furthermore, where appropriate, the cranial nerve palsy is grade 3, which occurs in the subject at a rate of about 1.1%; (2) the time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject ranges from about 17 days to about 60 days, and furthermore, where applicable, the median time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject is about 21 days; (3) the cranial nerve palsy affects cranial nerve III, V or VII; (4) the duration of cranial nerve palsy ranges from about 15 days to about 262 days, with the median duration of cranial nerve palsy being about 77 days; or (5) the treatment includes a corticosteroid; (d) CAR T-cell neurotoxicity is a peripheral neuropathy, with peripheral neuropathy occurring in the subject at a rate of approximately 2.8%, if applicable; or (e) CAR-T cell neurotoxicity is an adverse event induced by exercise and neurocognitive treatment, wherein, where appropriate, the adverse event induced by exercise and neurocognitive treatment is grade 1, and furthermore, where, where appropriate, the grade 1 adverse event induced by neurocognitive treatment and the subject's adverse event induced by neurocognitive treatment occur at a rate of approximately 0.6%. [64] The dose of T cells for use according to any one of claims 42-63, wherein the subject has one or more high-risk cytogenetic abnormalities selected from a group comprising Gain / AMP(1q), del(17p), t(4;14), t(14;16) or any combination thereof. [65] The dose of T cells for use according to claim 64, wherein the subject has at least two cytogenetic abnormalities and wherein the subject optionally has two, three, four, five or more cytogenetic abnormalities. [66] The dose of T cells for use according to claim 64 or 65, wherein the cytogenetic abnormality is a standard risk cytogenetic abnormality. [67] The dose of T cells for use according to any one of claims 1-66, wherein the administration of the dose of T cells is effective in obtaining a greater very good partial response (VGPR) or better in the subject compared to the administration of DPd or PVd treatment. [68] The dose of T cells for use according to claim 67, wherein the VGPR or better after administration of the treatment is about 81.3%. [69] The dose of T cells for use according to claim 67 or 68, wherein the VGPR or better after administration of DPd or PVd is about 45.5%. [70] The dose of T cells for use according to one of claims 67-69, wherein the treatment is more effective in obtaining a stringent complete response (sCR) in the subject compared to the administration of a DPd or PVd treatment. [71] The dose of T cells for use according to claim 70, wherein the sCR after administration of the treatment is about 58.2%. [72] The dose of T cells for use according to claim 70 or 71, wherein the sCR after administration of DPd or PVd is about 15.2%. [73] The dose of T cells for use according to any one of claims 67-72, wherein the administration of the dose of T cells reduces the risk of disease progression or death in the subject. [74] The dose of T cells for use according to claim 73, wherein the risk of disease progression or death is reduced compared to treatment with daratumumab pomalidomide dexamethasone (DPd) or pomalidomide bortezomib dexamethasone (PVd). [75] The dose of T cells for use according to claim 73, wherein the risk of disease progression or death is reduced compared to administering a double-cell treatment. [76] The dose of T cells for use according to any one of claims 67-75, wherein the subject has a reduced risk of disease progression or death of about 60% to about 75%. [77] The dose of T cells for use according to claim 76, wherein the subject has an approximately 74% reduced risk of disease progression or death. [78] The dose of T cells for use according to any one of claims 67-77, wherein the IMiD is lenalidomide. [79] The dose of T cells for use according to one of claims 67-78, wherein the subject has received three or fewer prior lines of therapy. [80] The dose of T cells for use according to any one of claims 67-79, wherein the subject has received two or fewer prior lines of therapy. [81] The dose of T cells for use according to any one of claims 67-80, wherein the subject has received only one prior line of therapy. [82] The dose of T cells for use according to any one of claims 67-81, wherein the method is effective in obtaining an overall response rate (ORR) of about 75% to about 100%. [83] The dose of T cells for use according to claim 82, wherein the ORR is about 84.6%. [84] The dose of T cells for use according to any one of claims 67-83, wherein the treatment is effective in maintaining the median progression-free survival (PFS) of the subject compared to administration of DPd or PVd treatment. [85] The dose of T cells for use according to claim 84, wherein the PFS after 12 months following administration of the treatment is approximately 75.9%. [86] The dose of T cells for use according to claim 84 or 85, wherein the PFS 12 months after administration of DPd or PVd is about 48.6%. [87] The dose of T cells for use according to any one of claims 67-86, wherein the method further comprises treating the subject against a CAR-T-associated adverse event after administration of the dose of T cells, wherein the method optionally comprises administering a treatment to the subject to alleviate the CAR-T-associated adverse event, wherein optionally the CAR-T-associated adverse event comprises cytokine release syndrome (CRS) and / or neurotoxicity, wherein optionally: (a) the CAR-T-associated adverse event is a CRS, wherein the CRS may further occur in the subject at a rate of approximately 60% to approximately 90% or at a rate of approximately 76.1%, wherein the maximum toxicity grade of the CRS may further be Grade 1, Grade 2 or Grade 3, wherein further: (1) the maximum toxicity level of the CRS is Grade 1, where, where applicable, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 52.8%; (2) the maximum toxicity level of the CRS is grade 2, where, where applicable, the maximum toxicity level of grade 2 occurs in the subject at a rate of approximately 22.2%; (3) the maximum toxicity level of the CRS is grade 3, where, where applicable, the maximum toxicity level of grade 3 occurs in the subject at a rate of approximately 1.1%; (4) the time to first onset of CRS is in the range of about 1 to about 23 days, with a median time to first onset of CRS of about 8 days where applicable; (5) the duration of the CRS ranges from about 1 to about 17 days, with a median duration of about 3 days where applicable; or (6) the treatment includes tocilizumab, oxygen, a corticosteroid, a vasopressor or any combination thereof; or (b) the CAR-T-associated adverse event is neurotoxicity, where appropriate the neurotoxicity includes an immune effector cell-associated neurotoxicity syndrome or associated symptom, movement and neurocognitive neurotoxicity, standard treatment-arm adverse event of neurotoxicity, a non-immune effector cell-associated neurotoxicity syndrome or associated symptom, or any combination thereof, where appropriate the neurotoxicity is an immune effector cell-associated neurotoxicity syndrome or associated neurotoxicity: (1) The immune effector cell-associated neurotoxicity syndrome or associated symptom occurs in the subject at a rate of approximately 4.5%; (2) the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1 or Grade 2, wherein the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 1, furthermore, where appropriate, the maximum toxicity level of Grade 1 occurs in the subject at a rate of approximately 3.4%, or where, where appropriate, the maximum toxicity level of the immune effector cell-associated neurotoxicity syndrome or associated symptom is Grade 2, furthermore, where appropriate, the maximum toxicity level of Grade 2 occurs in the subject at a rate of approximately 1.1%; (3) the time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 6 to about 15 days, with a median time to onset of immune effector cell-associated neurotoxicity syndrome or associated symptom of about 9.5 days, if applicable; (4) the duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom ranges from about 1 to about 6 days, with the median duration of the immune effector cell-associated neurotoxicity syndrome or associated symptom being about 2 days; or (5) the treatment includes a corticosteroid and / or tocilizumab. [88] The dose of T cells for use according to claim 87, wherein the neurotoxicity is CAR-T cell neurotoxicity, wherein optionally the CAR-T cell neurotoxicity occurs in the subject at a rate of about 17.0%, wherein optionally the CAR-T cell neurotoxicity comprises grade 3 / 4 neurotoxicity, grade 5 neurotoxicity, cranial nerve palsy, peripheral neuropathy, a movement event and a neurocognitive treatment-related adverse event or any combination thereof, further optionally comprising: (a) CAR-T cell neurotoxicity is grade 3 / 4 neurotoxicity, occurring in approximately 2.3% of the subject; (b) CAR-T cell neurotoxicity is grade 5 neurotoxicity; (c) CAR-T cell neurotoxicity is cranial nerve palsy, with cranial nerve palsy occurring in the subject at a rate of approximately 9.1%, where: (1) the cranial nerve palsy is grade 2 or grade 3, where furthermore, where appropriate, the cranial nerve palsy is grade 2, which occurs in the subject at a rate of about 8.0%, and where furthermore, where appropriate, the cranial nerve palsy is grade 3, which occurs in the subject at a rate of about 1.1%; (2) the time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject ranges from about 17 days to about 60 days, and furthermore, where applicable, the median time to onset of cranial nerve paralysis after administration of the dose of T cells to the subject is about 21 days; (3) the cranial nerve palsy affects cranial nerve III, V or VII; (4) the duration of cranial nerve palsy ranges from about 15 days to about 262 days, with the median duration of cranial nerve palsy being about 77 days; or (5) the treatment includes a corticosteroid; (d) CAR T-cell neurotoxicity is a peripheral neuropathy, with peripheral neuropathy occurring in the subject at a rate of approximately 2.8%, if applicable; or (e) CAR-T cell neurotoxicity is an adverse event induced by exercise and neurocognitive treatment, wherein, where appropriate, the adverse event induced by exercise and neurocognitive treatment is grade 1, and furthermore, where, where appropriate, the grade 1 adverse event induced by neurocognitive treatment and the subject's adverse event induced by neurocognitive treatment occur at a rate of approximately 0.6%. [89] The dose of T cells for use according to any one of claims 67-88, wherein the subject has one or more high-risk cytogenetic abnormalities selected from a group comprising Gain / AMP(1q), del(17p), t(4;14), t(14;16) or any combination thereof. [90] The dose of T cells for use according to claim 89, wherein the subject has at least two cytogenetic abnormalities and wherein the subject optionally has two, three, four, five or more cytogenetic abnormalities. [91] The dose of T cells for use according to claim 89 or 90, wherein the cytogenetic abnormality is a standard risk cytogenetic abnormality.

Images