Treatment of Multiple Myeloma

Abstract

Provided herein is a method for treating multiple myeloma, comprising administering an inhibitor of nuclear receptor-associated SET domain protein 2 (NSD2) to a subject with multiple myeloma. Also provided is a method of use when the multiple myeloma has previously been determined to have a 4:14 chromosomal translocation (t(4;14)).

Description

[Technical field]

[0001] I. CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 154,104, filed February 26, 2021, which is incorporated by reference in its entirety for all purposes.

[0002] II. Field Provided herein is a method of treating multiple myeloma, comprising administering an inhibitor of nuclear receptor-associated SET domain protein 2 (NSD2) to a subject with multiple myeloma.Also provided is a method of treating multiple myeloma where the multiple myeloma has previously been determined to have a 4:14 chromosomal translocation (t(4;14)). [Background technology]

[0003] III. Background Multiple myeloma (MM) is a cancer of plasma cells in the bone marrow. Normally, plasma cells produce antibodies and play a key role in immune function. However, uncontrolled proliferation of these cells leads to bone pain and fractures, anemia, infections, and other complications. Multiple myeloma is the second most common hematologic malignancy, but the exact cause of multiple myeloma remains unknown. Multiple myeloma causes high levels of proteins, including but not limited to M protein and other immunoglobulins (antibodies), albumin, and beta-2-microglobulin, in the blood, urine, and organs, except in some patients (estimated at 1%-5%) whose myeloma cells do not secrete these proteins (called nonsecretory myeloma). M protein, short for monoclonal protein, also known as paraprotein, is a particularly abnormal protein produced by myeloma plasma cells and can be found in the blood or urine of almost all multiple myeloma patients, except those with nonsecretory myeloma or whose myeloma cells produce immunoglobulin light chains along with heavy chains.

[0004] Skeletal symptoms, including bone pain, are among the most clinically significant symptoms of multiple myeloma. Malignant plasma cells release osteoclast stimulating factors (including IL-1, IL-6, and TNF) that leach calcium from bone, causing lytic lesions; hypercalcemia is another symptom. Osteoclast stimulating factors, also called cytokines, can prevent myeloma cells from undergoing apoptosis, or dying. Fifty percent of patients have radiologically detectable myeloma-associated skeletal lesions at the time of diagnosis. Other common clinical manifestations of multiple myeloma include polyneuropathy, anemia, hyperviscosity, infections, and renal insufficiency.

[0005] Cytogenetic features are important prognostic markers in multiple myeloma. Nearly 15 percent of newly diagnosed MM patients harboring the t(4;14) chromosomal translocation exhibit poor prognosis, including shorter progression-free survival (PFS) and overall survival (OS), which are only partially mitigated by existing therapies. Thus, there is a great need for therapeutic strategies specific to this subpopulation. Ortiz et al., Blood (2019) 134 (Supplement_1): 366. Summary of the Invention

[0006] IV.Overview Provided herein is a method of treating multiple myeloma, the method comprising administering to a subject with multiple myeloma an inhibitor of nuclear receptor-binding SET domain protein 2 (NSD2).

[0007] Also provided are methods of use where the multiple myeloma has previously been determined to have a 4:14 chromosomal translocation (t(4;14)).

[0008] The following non-limiting embodiments are provided.

[0009] Embodiment 1 is a method of treating multiple myeloma, comprising administering to a subject with multiple myeloma a therapeutically effective amount of an inhibitor of nuclear receptor-binding SET domain protein 2 (NSD2).

[0010] Embodiment 2 is the method of embodiment 1, wherein the multiple myeloma has previously been determined to have a 4:14 chromosomal translocation (t(4;14)).

[0011] Embodiment 3 is the method of embodiment 2, wherein the t(4;14) results in a disruption of the NSD2 gene.

[0012] Embodiment 4 is the method of embodiment 3, wherein the disruption of the NSD2 gene is located after the transcription start site of NSD2.

[0013] Embodiment 5 is the method of embodiment 3 or embodiment 4, wherein the disruption of the NSD2 gene is located after the translation start site of NSD2.

[0014] Embodiment 6 is a method according to any one of embodiments 3 to 5, wherein the disruption of the NSD2 gene is located before the translation stop site of NSD2.

[0015] Embodiment 7 is a method according to any one of embodiments 3 to 5, wherein the disruption of the NSD2 gene is located before the first coding exon of the NSD2 gene.

[0016] Embodiment 8 is the method of any one of embodiments 3 to 6, wherein the disruption of the NSD2 gene is located in the first coding exon of the NSD2 gene.

[0017] Embodiment 9 is a method according to any one of embodiments 3 to 6, wherein the disruption of the NSD2 gene is located between the start of the first coding exon and the start of the second coding exon of the NSD2 gene.

[0018] Embodiment 10 is the method of any one of embodiments 3 to 9, wherein the disruption of the NSD2 gene is located at or after genomic position 1,871,393 of the Genome Reference Consortium Human Build 38 Patch Release 13 (GRCh38.p13).

[0019] Embodiment 11 is a method according to any one of embodiments 3 to 10, wherein the disruption of the NSD2 gene is located between genomic position 1,871,393 and genomic position 1,900,655 of GRCh38.p13.

[0020] Embodiment 12 is a method according to any one of embodiments 3 to 10, wherein the disruption of the NSD2 gene is located at or after genomic position 1,900,655 of GRCh38.p13.

[0021] Embodiment 13 is a method according to any one of embodiments 3 to 10, wherein the t(4;14) is located between genomic position 1,900,655 and genomic position 1,982,207 of GRCh38.p13.

[0022] Embodiment 14 is the method of any one of embodiments 1 to 13, wherein the multiple myeloma expresses a truncated NSD2 protein.

[0023] Embodiment 15 is the method of any one of embodiments 1 to 13, wherein the multiple myeloma expresses full-length NSD2 protein.

[0024] Embodiment 16 is the method of embodiment 15, wherein the multiple myeloma expresses high levels of full-length NSD2 protein.

[0025] Embodiment 17 is the method of any one of embodiments 2 to 16, wherein the 4:14 chromosomal translocation (t(4;14)) has been identified by a method including in situ hybridization, PCR, RT-PCR, RNA sequencing, fluorescent in situ hybridization (FISH), transcript in situ hybridization, whole genome sequencing, whole exome sequencing, mixed ligation probe assay, mass spectrometry, and / or MALDI-TOF.

[0026] Embodiment 18 is a method according to any one of embodiments 1 to 17, wherein the inhibitor of NSD2 is selected from an antibody, a small molecule, an aptamer, an siRNA, and an antisense oligonucleotide.

[0027] Embodiment 19 is the method of any one of embodiments 1 to 18, comprising administering at least one second therapeutic agent.

[0028] Embodiment 20 is the method of embodiment 19, wherein the at least one second therapeutic agent is selected from a chemotherapeutic agent, a steroid, an immunomodulatory agent, a proteasome inhibitor, a histone deacetylase inhibitor, an anti-CD38 antibody, an anti-SLAMF7 antibody, an antibody drug conjugate, and a nuclear export inhibitor.

[0029] Embodiment 21 is the method of embodiment 19, wherein the at least one second therapeutic agent is selected from lenalidomide, thalidomide, pomalidomide, bortezomib, carfilzomib, ixazomib, panobinostat, melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, and bendamustine, dexamethasone, prednisone, daratumumab, isatuximab, elotuzumab, belantamab mafodotin-blmf, selinexor, pamidronate, zoledronic acid, and denosumab.

[0030] Embodiment 22 is an embodiment in which the at least one second therapeutic agent is: a) lenalidomide; b) Iberdomide; c) (S)-4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)methyl)benzyl)piperazin-1-yl)-3-fluorobenzonitrile; d) (i) lenalidomide, pomalidomide, or thalidomide; and (ii) dexamethasone; e) (i) carfilzomib, ixazomib, or bortezomib; (ii) lenalidomide; and (iii) dexamethasone; f) (i) bortezomib or carfilzomib; (ii) cyclophosphamide; and (iii) dexamethasone; g) (i) elotuzumab or daratumumab; (ii) lenalidomide; and (iii) dexamethasone; h) bortezomib, liposomal doxorubicin, and dexamethasone; i) panobinostat, bortezomib, and dexamethasone; j) elotuzumab, bortezomib, and dexamethasone; k) melphalan and prednisone with or without thalidomide or bortezomib; l) Vincristine, doxorubicin, and dexamethasone; m) dexamethasone, cyclophosphamide, etoposide, and cisplatin; and n) Dexamethasone, thalidomide, cisplatin, doxorubicin, cyclophosphamide, and etoposide, with or without bortezomib 20. The method of embodiment 19, wherein the method is selected from the group consisting of:

[0031] Embodiment 23 is a method of selecting a subject having multiple myeloma for treatment with an NSD2 inhibitor, comprising a step of determining whether the subject has a 4:14 chromosomal translocation (t(4;14)), and if the subject has t(4;14), selecting the subject for treatment with an NSD2 inhibitor.

[0032] Embodiment 24 is a method of predicting whether a subject having multiple myeloma will benefit from treatment with an NSD2 inhibitor, comprising a step of determining whether the subject has a 4:14 chromosomal translocation (t(4;14)), and if the subject has the t(4;14) translocation, the subject is predicted to benefit from treatment with an NSD2 inhibitor.

[0033] Embodiment 25 is a method according to embodiment 23 or embodiment 24, wherein the t(4;14) results in a disruption of the NSD2 gene.

[0034] Embodiment 26 is the method of embodiment 25, wherein the disruption of the NSD2 gene is located after the transcription start site of NSD2.

[0035] Embodiment 27 is the method of embodiment 25 or embodiment 26, wherein the disruption of the NSD2 gene is located after the translation start site of NSD2.

[0036] Embodiment 28 is a method according to any one of embodiments 25 to 27, wherein the disruption of the NSD2 gene is located before the translation stop site of NSD2.

[0037] Embodiment 29 is a method according to any one of embodiments 25 to 28, wherein the disruption of the NSD2 gene is located before the first coding exon of the NSD2 gene.

[0038] Embodiment 30 is a method according to any one of embodiments 25 to 28, wherein the disruption of the NSD2 gene is located in the first coding exon of the NSD2 gene.

[0039] Embodiment 31 is a method according to any one of embodiments 25 to 28, wherein the disruption of the NSD2 gene is located between the start of the first coding exon and the start of the second coding exon of the NSD2 gene.

[0040] Embodiment 32 is the method of any one of embodiments 25 to 31, wherein the disruption of the NSD2 gene is located at or after genomic position 1,871,393 of Genome Reference Consortium Human Build 38 Patch Release 13 (GRCh38.p13).

[0041] Embodiment 33 is a method according to any one of embodiments 25 to 32, wherein the disruption of the NSD2 gene is located between genomic position 1,871,393 and genomic position 1,900,655 of GRCh38.p13.

[0042] Embodiment 34 is a method according to any one of embodiments 25 to 32, wherein the disruption of the NSD2 gene is located at or after genomic position 1,900,655 of GRCh38.p13.

[0043] Embodiment 35 is a method according to any one of embodiments 25 to 32, wherein the t(4;14) is located between genomic position 1,900,655 and genomic position 1,982,207 of GRCh38.p13.

[0044] Embodiment 36 is a method according to any one of embodiments 23 to 35, wherein the multiple myeloma expresses a truncated NSD2 protein.

[0045] Embodiment 37 is a method according to any one of embodiments 23 to 35, wherein the multiple myeloma expresses full-length NSD2 protein.

[0046] Embodiment 38 is the method of embodiment 37, wherein the multiple myeloma expresses high levels of full-length NSD2 protein.

[0047] Embodiment 39 is the method of any one of embodiments 23 to 38, wherein determining whether the subject has t(4;14) comprises in situ hybridization, PCR, RT-PCR, RNA sequencing, fluorescent in situ hybridization (FISH), transcript in situ hybridization, whole genome sequencing, whole exome sequencing, mixed ligation probe assay, mass spectrometry, and / or MALDI-TOF.

[0048] Embodiment 40 is a method according to any one of embodiments 23 to 39, further comprising administering an NSD2 inhibitor.

[0049] Embodiment 41 is the method of embodiment 40, wherein the inhibitor of NSD2 is selected from an antibody, a small molecule, an aptamer, an siRNA, and an antisense oligonucleotide.

[0050] Embodiment 42 is the method of embodiment 40 or embodiment 41, comprising administering at least one second therapeutic agent.

[0051] Embodiment 43 is the method of embodiment 42, wherein the at least one second therapeutic agent is selected from a chemotherapeutic agent, a steroid, an immunomodulatory agent, a proteasome inhibitor, a histone deacetylase inhibitor, an anti-CD38 antibody, an anti-SLAMF7 antibody, an antibody drug conjugate, and a nuclear export inhibitor.

[0052] Embodiment 44 is the method of embodiment 42, wherein the at least one second therapeutic agent is selected from lenalidomide, thalidomide, pomalidomide, bortezomib, carfilzomib, ixazomib, panobinostat, melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, bendamustine, dexamethasone, prednisone, daratumumab, isatuximab, elotuzumab, belantamab mafodotin-blmf, selinexor, pamidronate, zoledronic acid, and denosumab.

[0053] Embodiment 45 is an embodiment in which the at least one second therapeutic agent is: a) lenalidomide; b) Iberdomide; c) (S)-4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)methyl)benzyl)piperazin-1-yl)-3-fluorobenzonitrile; d) (i) lenalidomide, pomalidomide, or thalidomide; and (ii) dexamethasone; e) (i) carfilzomib, ixazomib, or bortezomib; (ii) lenalidomide; and (iii) dexamethasone; f) (i) bortezomib or carfilzomib; (ii) cyclophosphamide; and (iii) dexamethasone; g) (i) elotuzumab or daratumumab; (ii) lenalidomide; and (iii) dexamethasone; h) bortezomib, liposomal doxorubicin, and dexamethasone; i) panobinostat, bortezomib, and dexamethasone; j) elotuzumab, bortezomib, and dexamethasone; k) melphalan and prednisone with or without thalidomide or bortezomib; l) Vincristine, doxorubicin, and dexamethasone; m) dexamethasone, cyclophosphamide, etoposide, and cisplatin; and n) Dexamethasone, thalidomide, cisplatin, doxorubicin, cyclophosphamide, and etoposide, with or without bortezomib 43. The method of embodiment 42, wherein the

[0054] V. Brief description of the drawings [Brief description of the drawings]

[0055] [Figure 1A] Figure 1A shows survival probability over time for 329 patients with newly diagnosed multiple myeloma (NDMM). [Figure 1B] FIG. 1B shows a density plot of overall survival for 329 NDMM patients. [Figure 1C] FIG. 1C shows the identification and grouping by patients with an overall survival of less than 24 months. [Figure 2A] FIG. 2A shows that translocations that disrupt the coding for the NSD2 protein correlate with shorter overall survival. [Figure 2B] FIG. 2B shows that patients were classified into three groups: no destruction, early destruction, and late destruction, with each group having an increasingly poor prognosis in terms of overall survival. [Figure 3A]Figure 3A shows that analysis of RNA-seq data identified three patient groups: (1) a group of full-length coding fusion transcripts with few or no DNA breaks, (2) a group of truncated fusion transcripts characterized by late DNA breaks, and (3) a small subset of patients in which fusion transcript expression was not measured. [Figure 3B] FIG. 3B shows that patients expressing the truncated fusion protein have inferior overall survival than patients expressing the full-length coding fusion transcript or patients not expressing the fusion transcript. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0056] VI. DETAILED DESCRIPTION OF THE PRESENTINVENTION A.Definition Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.All patents, applications, published applications and other publications are incorporated by reference in their entirety.If there are multiple definitions for a term in this specification, the definition in this section shall prevail unless otherwise stated.

[0057] As used herein, and in the appended claims, the indefinite articles "a" and "an" and the definite article "the" include plural referents as well as singular referents, unless the context clearly dictates otherwise.

[0058] As used herein, the terms "comprising" and "including" can be used synonymously. The terms "comprising" and "including" should be interpreted to specify the presence of the described features or components as referred to, but do not exclude the presence or addition of one or more features or components or groups thereof. Furthermore, the terms "comprising" and "including" are intended to include examples encompassed by the term "consisting of". As a result, the term "consisting of" can be used in place of the terms "comprising" and "including" to provide more specific embodiments of the present invention.

[0059] The term "consisting of" means that the subject matter has at least 90%, 95%, 97%, 98% or 99% of the recited features or components that make it up. In another embodiment, the term "consisting of" excludes from the scope of any succeeding recitation any other features or components, excepting those that are not essential to the technical effect to be achieved.

[0060] As used herein, the term "or" should be interpreted as an inclusive "or" meaning any one or any combination. Thus, "A, B or C" means any of the following: "A; B; C; A and B; A and C; B and C; A, B and C." Exceptions to this definition occur only when combinations of elements, features, steps or acts are in some way inherently mutually exclusive.

[0061] As used herein, unless otherwise specified, the terms "about" and "approximately," when used in conjunction with a dose, amount, or weight percentage of a component of a composition or dosage form, refer to a dose, amount, or weight percentage that is recognized by those skilled in the art to provide an equivalent pharmacological effect to that obtained from the specified dose, amount, or weight percentage. In certain embodiments, the terms "about" and "approximately," when used in this context, contemplate a dose, amount, or weight percentage that is within 30%, within 20%, within 15%, within 10%, or within 5% of the specified dose, amount, or weight percentage.

[0062] As used herein, the term "Genome Reference Consortium human build 38 patch release 13," abbreviated as "GRCh38.p13" or "hg38," refers to the human reference genome assembly of that name dated February 28, 2019. The GenBank accession number for GRCh38 is GCA_000001405.28 and is also available at RefSeq under accession number GCF_000001405.39.

[0063] As used herein, the terms "NSD2" and "nuclear receptor-associated SET domain protein 2" are used interchangeably to refer to an NSD2 polypeptide or a nucleic acid, such as a gene, encoding the polypeptide. The NSD2 polypeptide is a lysine histone methyltransferase (HMTase) (EC2.1.1.357) that specifically dimethylates lysine 36 of nucleosomal histone H3 and is involved in chromatin binding and gene transcription regulation in various biological processes.

[0064] An exemplary amino acid sequence of a human NSD2 polypeptide found in UniProt ID O96028 is as follows: MEFSIKQSPL SVQSVVKCIK MKQAPEILGS ANGKTPSCEV NRECSVFLSK AQLSSSLQEG 60 VMQKFNGHDA LPFIPADKLK DLTSRVFNGE PGAHDAKLRF ESQEMKGIGT PPNTTPIKNG 120 SPEIKLKITK TYMNGKPLFE SSICGDSAAD VSQSEENGQK PENKARRNRK RSIKYDSLLE 180 QGLVEAALVS KISSPSDKKI PAKKESCPNT GRDKDHLLKY NVGDLVWSKV SGYPWWPCMV 240 SADPLLHSYT KLKGQKKSAR QYHVQFFGDA PERAWIFEKS LVAFEGEGQF EKLCQESAKQ 300 APTKAEKIKL LKPISGKLRA QWEMGIVQAE EAASMSVEER KAKFTFLYVG DQLHLNPQVA 360 KEAGIAAESL GEMAESSGVS EEAAENPKSV REECIPMKRR RRACLCSSAE TLESHPDIGK 420 STPQKTAEAD PRRGVGSPPG RKKTTVSMPR SRKGDAASQF LVFCQKHRDE VVAEHPDASG 480 EEIEELLRSQ WSLLSEKQRA RYNTKFALVA PVQAEEDSGN VNGKKRNHTK RIQDPTEDAE 540 AEDTPRKLR TDKHSLRRKRD TITDKTARTS SYKAMEAASS LKSQAATKNL SDACKPLKKR 600 NRASTAASSA LGFSKSSSPS ASLTENEVSD SPGDEPSESP YESADETQTE VSVSSKKSER 660 GVTAKKEYVC QLCEKPGSLL LCEGPCCGAF HLACGLSRR PEGRFTCSEC ASGIHSCFVC 720 KESKTDVKRC VVTQCGKFYH EACVKKYPLT VFESRGFRCP LHSCVSCHAS NPSNRPSKG 780 KMMRCVRCPV AYHSGDACLA AGCSVIASNS IICTAHFTAR KGKRHHAHVN VSWCFVCSKG 840 GSLLCCESCP AAFHPDCLNI EMPDGSWFCN DCRAGKKLHF QDIIWVKLGN YRWWPAEVCH 900 PKNVPPNIQK MKHEIGEFPV FFFGSKDYYW THQARVFPYM EGDRGSRYQG VRGIGRVFKN 960 ALQEAEARFR EIKLQREARE TQESERKPPP YKHIKVNKPY GKVQIYTADI SEIPKCNCKP 1020 TDENPCGFDS ECLNRMLMFE CHPQVCPAGE FCQNQCFTKR QYPETKIIKT DGKGWGLVAK 1080 RDIRKGEFVN EYVGELIDEE ECMARIKHAH ENDITHFYML TIDKDRIIDA GPKGNYSRFM 1140 NHSCQPNCET LKWTVNGDTR VGLFAVCDIP AGTELTFNYN LDCLGNEKTV CRCGASNCSG 1200 FLGDRPKTST TLSSEEKGKK TKKKTRRRRA KGEGKRQSED ECFRCGDGGQ LVLCDRKFCT 1260 KAYHLSCLGL GKRPFGKWEC PWHHCDVCGK PSTSFCHLCP NSFCKEHQDG TAFSCTPDGR 1320 SYCCEHDLGA ASVRSTKTEK PPPEPGKPKG KRRRRRGWRR VTEGK 1365 (SEQ ID NO:1)

[0065] An exemplary human NSD2 gene sequence is available from Ensembl under the accession number ENSG00000109685.19. The NSD2 gene is located on the forward strand of chromosome 4 at nucleotides 1,871,393 to 1,982,207.

[0066] As used herein, unless otherwise specified, the terms "express" and "expression" refer to gene expression, including expression of nucleic acids (e.g., mRNA) and expression of polypeptides. Thus, "NSD2 expression" can be determined by evaluating the expression of NSD2 mRNA and / or the expression of NSD2 protein.

[0067] As used herein, unless otherwise specified, the term "treating" refers to the total or partial alleviation of a disorder, disease or condition, or one or more symptoms associated with a disorder, disease or condition, or slowing or halting the further progression or worsening of such symptoms, or alleviating or eradicating the cause of the disorder, disease or condition itself.

[0068] As used herein, unless otherwise specified, the term "prevent" refers to a method of delaying and / or arresting the onset, recurrence or spread of a disorder, disease or condition, in whole or in part; barring a subject from acquiring a disorder, disease, or condition; or reducing the risk that a subject will acquire a disorder, disease, or condition.

[0069] As used herein, unless otherwise specified, the term "managing" includes preventing the recurrence of a particular disease or disorder in a patient afflicted with it, extending the time that a patient afflicted with a disease or disorder remains in remission, reducing patient mortality, and / or maintaining a reduction in the severity or avoidance of symptoms associated with the disease or condition being managed.

[0070] As used herein, unless otherwise specified, the term "effective amount" or "therapeutically effective amount" in reference to a compound means an amount capable of treating, preventing, or managing a disorder, disease or condition, or a symptom thereof.

[0071] As used herein, unless otherwise specified, the term "subject" or "patient" includes animals, and in some embodiments, mammals. In some embodiments, the subject or patient is a human.

[0072] As used herein, unless otherwise specified, the term "sample" refers to a sample obtained from a subject. A sample may be derived from any biological tissue or fluid. In some embodiments, a sample is derived from a human, e.g., a subject or patient, e.g., a cancer patient, e.g., a multiple myeloma patient. A sample may include tissue, tissue section, cell, fluid, or extract thereof, and may be isolated by any means, e.g., from blood, serum, biopsy, lymph node biopsy, bone marrow biopsy, needle biopsy, aspirate, etc.

[0073] As used herein, unless otherwise specified, the term "recurrent" refers to a disorder, disease, or condition that has responded to treatment (e.g., achieved a partial or complete response) and then progressed. Treatment may include one or more lines of treatment. In some embodiments, the disorder, disease, or condition has been previously treated with one or more lines of treatment. In other embodiments, the disorder, disease, or condition has been previously treated with one, two, three, or four lines of treatment. In some embodiments, the disorder, disease, or condition is a hematological malignancy.

[0074] As used herein, unless otherwise specified, the term "refractory" refers to a disorder, disease, or condition that has not responded to previous treatment. In some embodiments, the disorder, disease, or condition has been previously treated with one, two, three, or four lines of treatment. In some embodiments, the disorder, disease, or condition has been previously treated with two or more lines of treatment and has not responded to the most recent treatment. In some embodiments, the disorder, disease, or condition is a hematological malignancy, particularly multiple myeloma.

[0075] In the context of cancer, e.g., hematological malignancies, inhibition can be assessed by, among others, inhibition of disease progression, inhibition of tumor growth, shrinkage of primary tumor, alleviation of tumor-related symptoms, inhibition of tumor secreted factors, delay in the appearance of primary or secondary tumors, slowing down the development of primary or secondary tumors, reducing the occurrence of primary or secondary tumors, slowing down or reducing the severity of secondary effects of disease, halting tumor growth and tumor regression, increasing time to progression (TTP), increasing progression-free survival (PFS), and increasing overall survival (OS). OS, as used herein, refers to the time from the start of treatment to death from any cause. TTP, as used herein, refers to the time from the start of treatment to tumor progression; TTP does not include death. In some embodiments, PFS refers to the time from the start of treatment to tumor progression or death. In some embodiments, PFS refers to the time from the first dose of compound to the first occurrence of disease progression or death from any cause. In some embodiments, the PFS rate is calculated using Kaplan-Meier estimates. Event-free survival (EFS) refers to the time from initiation of treatment to any treatment failure, including disease progression, treatment discontinuation for any reason, or death. In some embodiments, overall response rate (ORR) refers to the percentage of patients who achieve a response. In some embodiments, ORR refers to the sum of the percentage of patients who achieve a complete response and a partial response. In some embodiments, ORR refers to the percentage of patients whose best response is >= partial response (PR). In some embodiments, duration of response (DoR) is the time from achievement of response to relapse or disease progression. In some embodiments, DoR is the time from achievement of response >= partial response (PR) to relapse or disease progression. In some embodiments, DoR is the time from first documented response to first documented progressive disease or death. In some embodiments, DoR is the time from first documented response >= partial response (PR) to first documented progressive disease or death. In some embodiments, time to response (TTR) refers to the time from the first dose of a compound to the first recorded response.In some embodiments, TTR refers to the time from the first dose of compound to the first recorded response ≥ partial response (PR). In extreme cases, complete inhibition is referred to herein as prevention or chemoprevention. In this context, the term "prevention" includes either completely preventing the onset of clinically evident cancer or preventing the onset of preclinically evident stages of cancer. This definition is also intended to encompass the prevention of transformation into malignant cells, or arresting or reversing the progression of premalignant cells to malignant cells. This includes prophylactic treatment of those at risk of developing cancer.

[0076] As used herein, "multiple myeloma" refers to a hematological condition characterized by malignant plasma cells and includes the following disorders: monoclonal gammopathy of undetermined significance (MGUS); low-, intermediate-, and high-risk multiple myeloma; newly diagnosed multiple myeloma (including low-, intermediate-, and high-risk newly diagnosed multiple myeloma); transplant eligible and transplant ineligible multiple myeloma; smoldering (low-grade) multiple myeloma (including low-, intermediate-, and high-risk smoldering multiple myeloma); active multiple myeloma; solitary plasmacytoma; extramedullary plasmacytoma; plasma cell leukemia; central nervous system multiple myeloma; light chain myeloma; nonsecretory myeloma; immunoglobulin A myeloma; and immunoglobulin D myeloma; and immunoglobulin E myeloma; and multiple myeloma characterized by genetic abnormalities such as cyclin D translocations (e.g., t(11;14)(q13;q32); t(6;14)(p21;32); t(12;14)(p13;q32); or t(6;20);); MMSET translocations (e.g., t(4;14)(p16;q32)); MAF translocations (e.g., t(14;16)(q32;q32); t(20;22); t(16;22)(q11;q13); or t(14;20)(q32;q11)); or other chromosomal factors (e.g., deletions of chromosome 17p13 or chromosome 13; del(17 / 17p), non-hyperdiploidy, and gain(1q)). In some embodiments, the multiple myeloma is characterized by chromosomal translocation t(4;14). In some embodiments, the multiple myeloma is characterized according to the International Staging System for Multiple Myeloma (ISS). In some embodiments, the multiple myeloma is stage I multiple myeloma characterized by ISS (e.g., serum β2 microglobulin < 3.5 mg / L and serum albumin ≧ 3.5 g / dL). In some embodiments, the multiple myeloma is stage III multiple myeloma characterized by ISS (e.g., serum β2 microglobulin > 5.4 mg / L). In some embodiments, the multiple myeloma is stage II multiple myeloma characterized by ISS (e.g., neither stage I nor III).

[0077] In certain embodiments, treatment of multiple myeloma may be evaluated by the International Uniform Response Criteria for Multiple Myeloma (IURC) using the response and endpoint definitions provided below (see Durie BGM, Harousseau JL, Miguel JS, et al. International uniform response criteria for multiple myeloma. Leukemia, 2006; (10) 10: 1-7). [Table 1] Abbreviations: CR, complete response; FLC, free light chain; PR, partial response; SD, stable disease; sCR, stringent complete response; VGPR, very good partial response. a All response categories require two consecutive evaluations done at any time before the introduction of any new therapy; all categories do not require known evidence of progressive or new bone lesions if radiographs are performed. Radiographs are not required to meet these response requirements. b Confirmation by repeat bone marrow biopsy is not required. c The presence / absence of clonal cells is based on the kappa / lambda ratio. Analysis of abnormal kappa / lambda ratios by immunohistochemistry and / or immunofluorescence requires at least 100 plasma cells. Abnormal ratios reflecting the presence of abnormal clones are kappa / lambda >4:1 or <1:2. d Measurable disease defined by at least one of the following measurements: bone marrow plasma cells ≥ 30%; serum M protein ≥ 1 g / dl (≥ 10 gm / l) [10 g / l]; urinary M protein ≥ 200 mg / 24 hr; serum FLC assay: involved FLC level ≥ 10 mg / dl (≥ 100 mg / l); however, serum FLC ratio is abnormal.

[0078] As used herein, ECOG status refers to the Eastern Cooperative Oncology Group (ECOG) performance status as set forth below (Oken M, et al Toxicity and response criteria of the Eastern Cooperative Oncology Group. Am J Clin Oncol 1982;5(6):649-655). [Table 2]

[0079] In certain embodiments, stable disease or lack thereof can be determined by methods known in the art, such as assessment of patient symptoms, physical examination, visualization of the tumor imaged using, for example, FDG-PET (fluorodeoxyglucose positron emission tomography), PET / CT (positron emission tomography / computed tomography) scans, MRI (magnetic resonance imaging) of the brain and spine, CSF (cerebrospinal fluid), ophthalmic examination, vitreous fluid sampling, retinal photography, bone marrow evaluation, and other commonly accepted evaluation methods.

[0080] As used herein, unless otherwise specified, the terms "co-administration" and "in combination with" include administration of one or more therapeutic agents (e.g., a compound provided herein and another anti-cancer or supportive care agent) simultaneously, in parallel, or sequentially without specific time limitations. In some embodiments, the agents are present in a cell or in a patient's body at the same time or exert their biological or therapeutic effects at the same time. In some embodiments, the therapeutic agents are present in the same composition or unit dosage form. In other embodiments, the therapeutic agents are present in separate compositions or unit dosage forms.

[0081] The term "supportive care agent" refers to any substance that treats, prevents, or manages adverse effects from treatment with another therapeutic agent.

[0082] As used herein, "induction therapy" refers to the first treatment given for a disease or the first treatment given for the purpose of inducing complete remission in a disease such as cancer. When used by itself, induction therapy is recognized as the best available treatment. If residual cancer is detected, the patient is treated with another treatment, called reinduction. If the patient is in complete remission after induction therapy, additional consolidation and / or maintenance therapy is given to extend remission or potentially cure the patient.

[0083] As used herein, "consolidation therapy" refers to the treatment given for disease after remission is first achieved.For example, consolidation therapy for cancer is the treatment given after the cancer disappears after the first therapy.Consolidation therapy can include treatment with radiation therapy, stem cell transplantation or cancer chemotherapy.Consolidation therapy is also called intensive therapy and post-remission therapy.

[0084] As used herein, "maintenance therapy" refers to treatment given for a disease to prevent or delay recurrence after remission or optimal response has been achieved. Maintenance therapy may include chemotherapy, hormonal therapy, or targeted therapy.

[0085] "Remission" as used herein is the reduction or disappearance of signs and symptoms of cancer, such as multiple myeloma. In partial remission, some but not all signs and symptoms of cancer disappear. In complete remission, all signs and symptoms of cancer disappear, but cancer may still be present in the body.

[0086] As used herein, "transplantation" refers to high-dose therapy with stem cell rescue. Hematopoietic (blood) stem cells or bone marrow stem cells are used not as a treatment but to rescue patients after high-dose therapy, such as high-dose chemotherapy and / or radiation. Transplantation includes "autologous" stem cell transplantation (ASCT), which refers to the harvesting of a patient's own stem cells to be used as replacement cells. In some embodiments, transplantation also includes tandem or multiple transplantation.

[0087] The term "biologic therapy" refers to the administration of biological therapeutic agents such as umbilical cord blood, stem cells and growth factors.

[0088] B. Selection of treatment targets In some embodiments, provided herein is a method of treating multiple myeloma, the method comprising administering to a subject having multiple myeloma an inhibitor of nuclear receptor-binding SET domain protein 2 (NSD2).

[0089] In some embodiments, provided herein is a method for selecting a subject with multiple myeloma for treatment with NSD2 inhibitor, comprising determining whether the subject has 4:14 chromosomal translocation (t(4;14)).In some embodiments, if the subject has t(4;14), the subject is selected for treatment with NSD2 inhibitor.

[0090] In some embodiments, provided herein is a method for predicting whether a subject with multiple myeloma will benefit from treatment with an NSD2 inhibitor, comprising determining whether the subject has 4:14 chromosomal translocation (t(4;14)).In some embodiments, if the subject has t(4;14) translocation, the subject is predicted to benefit from treatment with an NSD2 inhibitor.

[0091] A subject can be identified as having multiple myeloma by any available method, hi some embodiments, the subject has previously been determined to have multiple myeloma.

[0092] In some embodiments, the cancer cells have a chromosomal rearrangement resulting in a t(4;14) chromosomal translocation. In some embodiments, the t(4;14) chromosomal translocation is in the NSD2 gene.

[0093] In some embodiments, the cancer cells have previously been determined to have the t(4;14) chromosomal translocation.

[0094] In some embodiments, the t(4;14) chromosomal translocation results in a disruption of the NSD2 gene.

[0095] In some embodiments, the t(4;14) chromosomal translocation does not result in a disruption of the protein coding region of the NSD2 gene.

[0096] In some embodiments, the t(4;14) chromosomal translocation leads to disruption of the protein coding region of the NSD2 gene. In some embodiments, translocations that disrupt the protein coding region of the NSD2 gene are correlated with shorter overall survival.

[0097] In some embodiments, multiple myeloma expresses truncated NSD2 protein.In some embodiments, multiple myeloma expresses fusion protein comprising truncated or full-length NSD2 protein.In some embodiments, patients who express truncated fusion protein comprising NSD2 have poorer overall survival than patients who express full-length coding fusion transcript or patients who do not express fusion transcript.

[0098] In some embodiments, the disruption of the NSD2 gene is located in the first coding exon of the NSD2 gene.

[0099] In some embodiments, the disruption of NSD2 gene is based on the position of translocation breakpoint and is a first half disruption.The first half disruption is a disruption between the transcription start site of NSD2 gene and the translation start site of NSD2 gene.In some embodiments, the first half disruption is between genomic position 1,871,393 and genomic position 1,900,655 of GRCh38.p13.

[0100] In some embodiments, the disruption of the NSD2 gene is a late disruption based on the position of the translocation breakpoint. In some embodiments, the late disruption is a disruption in the NSD2 gene after the translation start site. In some embodiments, the late disruption is a disruption downstream of genomic position 1,900,655 of GRCh38.p13. In some embodiments, the late disruption is a disruption between genomic position 1,900,655 and genomic position 1,983,934 of GRCh38.p13.

[0101] In some embodiments, the 4:14 chromosomal translocation is located before genomic position 1,871,393 in Genome Reference Consortium Human Build 38 Patch Release 13 ("GRCh38.p13"). In some such embodiments, there is no disruption in the NSD2 gene.

[0102] In some embodiments, t(4;14) is located after the transcription start site of NSD2 gene. In some embodiments, t(4;14) is located between the start of the first protein-coding exon and the start of the second protein-coding exon of NSD2 gene. In some embodiments, t(4;14) is located at or after the start of the second protein-coding exon of NSD2 gene.

[0103] In some embodiments, the t(4;14) is located at or after genomic position 1,871,393 of GRCh38.p13. In some embodiments, the t(4;14) is located between genomic position 1,871,393 and genomic position 1,900,655 of GRCh38.p13.

[0104] In some embodiments, the t(4;14) is located at or after genomic position 1,900,655 of GRCh38.p13. In some embodiments, the t(4;14) is located between genomic position 1,900,655 and genomic position 1,983,934 of GRCh38.p13.

[0105] In some embodiments, t(4;14) chromosomal translocation is identified directly or indirectly by analyzing NSD2 protein expression. In some embodiments, NSD2 protein expression is determined and compared to a reference (e.g., a reference sample, or a reference value, or any other comparison that indicates whether or not, or to what extent, and / or in what form the cancer expresses NSD2 polypeptide). In some embodiments, NSD2 expression in multiple myeloma is determined relative to NSD2 expression in non-cancerous cells. In some embodiments, analysis and quantification of NSD2 expression in patient samples, cells, and / or cell lines is determined by immunohistochemical and / or immunofluorescence techniques.

[0106] In some embodiments, the t(4;14) chromosomal translocation is identified by methods including polymerase chain reaction (PCR), reverse transcription PCR (RT-PCR including real-time RT-PCR, qRT-PCR), RNA sequencing (RNA-seq), in situ hybridization (ISH), fluorescent in situ hybridization (FISH), transcript in situ hybridization, whole genome sequencing (WGS), whole exome sequencing (WES), multiplex ligation-dependent probe assay (MLPA), mass spectrometry (MS), and / or matrix-assisted laser desorption / ionization time-of-flight MS (MALDI-TOF MS).

[0107] The term "polymerase chain reaction" or "PCR" as used herein refers to a procedure in which small amounts of nucleic acid, RNA and / or DNA, are amplified. Generally, sequence information from the ends or beyond the region of interest must be available so that oligonucleotide primers can be designed; these primers will be identical or similar in sequence to opposite strands of the template to be amplified. The 5' terminal nucleotides of the two primers may match the ends of the amplified material. PCR can be used to amplify specific RNA sequences, specific DNA sequences of total genomic DNA, and cDNA transcribed from total cellular RNA, bacteriophage or plasmid sequences, etc.

[0108] Other PCR-based methods can also be used. Examples of PCR methods can be found in the literature. Examples of PCR assays can be found, for example, in U.S. Patent No. 6,927,024. A non-limiting example of RT-PCR can be found in U.S. Patent No. 7,122,799. A non-limiting method of fluorescent in situ PCR is described in U.S. Patent No. 7,186,507.

[0109] In some embodiments, real-time reverse transcription PCR (qRT-PCR) can be used for both detection and quantification of RNA targets (Bustin, et al., 2005, Clin. Sci., 109:365-379). Quantitative results obtained by qRT-PCR are generally more informative than qualitative data. Thus, in some embodiments, qRT-PCR-based assays can be useful for measuring mRNA levels during cell-based assays. qRT-PCR methods are also useful for monitoring patient treatment. Examples of qRT-PCR-based methods can be found, for example, in U.S. Pat. No. 7,101,663.

[0110] In contrast to conventional reverse transcriptase-PCR and agarose gel analysis, real-time PCR provides quantitative results. An additional advantage of real-time PCR is that it is relatively easy and convenient to use. Instruments for real-time PCR, such as the Applied Biosystems 7500, are commercially available, as are reagents such as TaqMan Sequence Detection chemistry. For example, TaqMan® gene expression assays can be used according to the manufacturer's instructions. These kits are pre-formulated gene expression assays for rapid and reliable detection and quantification of human, mouse and rat mRNA transcripts. An exemplary PCR program is, for example, 40 cycles of 50°C for 2 minutes, 95°C for 10 minutes, 95°C for 15 seconds, then 60°C for 1 minute.

[0111] The term "RNA sequencing" or "RNA-Seq" typically refers to a method that includes the steps of (1) isolating RNA; (2) depleting ribosomal RNA; (3) synthesizing cDNA; and (4) sequencing the cDNA by next-generation sequencing (NGS). The technology of RNA-Seq is well known to those skilled in the art, and the method is described in Waern et al., Methods Mol Biol., 2011, 759: 125-132; Wilhelm et al., Nature Protocols, 2010, 5(2):255-66; and Hoeijmakers et al., Methods Mol Biol., 2013, 923:221-39.

[0112] The term "in situ hybridization" or "ISH" refers to a technique that can be used to determine mRNA levels (reviewed in A. K. Raap (1998) Mutat. Res. 400:287-298). In situ hybridization techniques allow for the visual detection of mRNA in cells by incubating the cells with a labeled (e.g., fluorescently or digoxigenin-labeled) oligonucleotide probe that hybridizes to the mRNA of interest and then examining the cells under a microscope.

[0113] For more precise gene mapping, fluorescent in situ hybridization (FISH) technology can be used. In particular, high-resolution FISH technology (A. Palotie et al. (1996) Ann. Med. 28:101-106) uses free chromatin, DNA fiber, or mechanically stretched chromosomes to map gene sequences ranging in size from a few kilobases to 300 kb. Alternatively, the chromosomal location of genes can be determined from a suitable genome database, for example, the Homo sapiens genome database available at the Entrez Genome Web site (National Center for Biotechnology Information, Bethesda, MD).

[0114] The term "whole genome sequencing" or "WGS," as used herein, refers to a method of sequencing an entire genome.

[0115] The term "whole exome sequencing" or "WES," as used herein, refers to a method of sequencing all protein-coding regions (exons) of the genome.

[0116] The term "mass spectrometry" or "mass spec" or "MS" as used herein refers to an analytical technique for measuring the mass-to-charge ratio of ions. This is accomplished by ionizing a sample to separate ions of different masses and recording their relative abundance by measuring the intensity of the ion flux. A typical mass spectrometer includes three parts: an ion source, a mass analyzer, and a detector system. The ion source is the part of the mass spectrometer that ionizes the substance under analysis (analyte). The ions are then transported by a magnetic or electric field to the mass analyzer where the ions are separated according to their mass-to-charge ratio (m / z). In many mass spectrometers, two or more mass analyzers are used in the case of tandem mass spectrometry (MS / MS). The detector records the charge induced or the current generated as the ions pass through or impact a surface. A mass spectrum is the result of measuring the signal generated by the detector as the mass analyzer scans the m / z ions. Exemplary mass spectrometry analytical techniques include electrospray ionization mass spectrometry (ESI), high resolution mass spectrometry (HRMS), liquid chromatography mass spectrometry (LCMS), and liquid chromatography tandem mass spectrometry (LC-MS / MS). Also exemplary is matrix-assisted laser desorption / ionization (MALDI) mass spectrometry, such as MALDI-TOF mass spectrometry, where MALDI is the ion source and the mass analyzer is a time-of-flight (TOF) mass spectrometer.

[0117] C. Inhibitors of nuclear receptor-associated SET domain protein 2 (NSD2) In some embodiments, the inhibitor of NSD2 is an antibody. The term "antibody" is used in the broadest sense herein and includes fully assembled antibodies, antibody fragments (e.g., Fab, F(ab')2, Fv, and other fragments) that retain the ability to specifically bind to an antigen, single chain antibodies, diabodies, antibody chimeras, hybrid antibodies, bispecific antibodies, and humanized antibodies.

[0118] In some embodiments, the inhibitor of NSD2 is a small molecule. The term "small molecule" is used in the broadest sense herein and includes molecules less than 1,000 daltons, including synthetic inorganic, organometallic, and organic molecules.

[0119] In some embodiments, the inhibitor of NSD2 is an aptamer. The term "aptamer" as used herein is an oligonucleotide or peptide molecule that specifically binds to a target. In some embodiments, the aptamer is an oligonucleotide having about 15 to about 100 nucleotides, such as about 15 to about 50 nucleotides.

[0120] In some embodiments, the inhibitor of NSD2 is a small interfering RNA (siRNA). The term "siRNA" or "small interfering RNA" or "short interfering RNA" or "silencing RNA" as used herein refers to a single- or double-stranded non-coding RNA of about 20 to about 27 bases or base pairs in length that interferes with the expression of a specific RNA with which it has a complementary nucleotide sequence. In some embodiments, the siRNA causes degradation of the mRNA and prevents translation of the protein product.

[0121] In some embodiments, the inhibitor of NSD2 is an antisense oligonucleotide. The term "antisense oligonucleotide" as used herein refers to a single-stranded oligonucleotide that is complementary to a specific sequence of a target gene or RNA and modulates its expression or splicing. In some embodiments, the antisense oligonucleotide is at least 10 nucleotides, such as at least 15 nucleotides, and may be about 15 to about 30 nucleotides, such as about 15 to about 25 nucleotides, and may contain one or more modifications compared to naturally occurring nucleotides.

[0122] In some embodiments, the specific amount of an inhibitor of NSD2 provided herein for use in the methods provided herein will be determined by factors such as the specific type of inhibitor used, the type of multiple myeloma being treated or managed, the severity and stage of the disease, the age, height, and / or weight of the subject being treated; and any optional additional active agents co-administered to the patient.

[0123] D.How to use In some embodiments, provided herein is a method of treating multiple myeloma, the method comprising administering to a subject having multiple myeloma an inhibitor of nuclear receptor-binding SET domain protein 2 (NSD2).

[0124] In some embodiments, provided herein is a method of treating multiple myeloma, the method comprising administering to a subject having multiple myeloma a therapeutically effective amount of an inhibitor of nuclear receptor-binding SET domain protein 2 (NSD2).

[0125] In some embodiments, the multiple myeloma is plasma cell leukemia (PCL).

[0126] In some embodiments, the multiple myeloma is newly diagnosed multiple myeloma.

[0127] In some embodiments, the multiple myeloma is relapsed or refractory. In some embodiments, the multiple myeloma is refractory to lenalidomide. In some embodiments, the multiple myeloma is refractory to pomalidomide. In some embodiments, the multiple myeloma is refractory to a combination of pomalidomide and a proteasome inhibitor. In some embodiments, the proteasome inhibitor is selected from bortezomib, carfilzomib, and ixazomib. In some embodiments, the multiple myeloma is refractory to a combination of pomalidomide and an inflammatory steroid. In some embodiments, the inflammatory steroid is selected from dexamethasone or prednisone. In some embodiments, the multiple myeloma is refractory to a combination of pomalidomide and a monoclonal antibody directed against CD38.

[0128] In some embodiments, provided herein is a method for achieving a complete response, partial response, or stable disease in a patient, the method comprising administering an inhibitor of NSD2 to a subject with multiple myeloma.

[0129] In some embodiments, also provided herein is a method for inducing a therapeutic response as assessed by the International Uniform Response Criteria (IURC) for multiple myeloma in a patient (see Durie BGM, Harousseau JL, Miguel JS, et al. International uniform response criteria for multiple myeloma. Leukemia, 2006; (10) 10: 1 7), comprising administering to a subject having multiple myeloma an inhibitor of NSD2.

[0130] In some embodiments, provided herein is a method for achieving a stringent complete response, a complete response, or a very good partial response in a patient as determined by International Uniform Response Criteria for Multiple Myeloma (IURC), comprising administering an inhibitor of NSD2 to a patient with multiple myeloma.

[0131] In some embodiments, provided herein is a method for achieving increased overall survival, progression-free survival, symptom-free survival, progression-free survival, or disease-free survival in a patient, the method comprising administering an inhibitor of NSD2 to a subject with multiple myeloma.

[0132] In some embodiments, also provided herein is a method of selecting a subject having multiple myeloma for treatment with an NSD2 inhibitor, the method comprising determining whether the subject has a 4:14 chromosomal translocation (t(4;14)), and if the subject has t(4;14), selecting the subject for treatment with an NSD2 inhibitor.

[0133] In some embodiments, there is provided a method of selecting a subject having multiple myeloma for treatment with an NSD2 inhibitor, comprising: a) obtaining a sample from a subject; b) determining whether the subject has a 4:14 chromosomal translocation (t(4;14)); c) if the subject has the t(4;14) chromosomal translocation, selecting the subject for treatment with an NSD2 inhibitor. Provided herein is a method comprising:

[0134] In some embodiments, provided herein is a method of predicting whether a subject having multiple myeloma will benefit from treatment with an NSD2 inhibitor, the method comprising determining whether the subject has a 4:14 chromosomal translocation (t(4;14)), and if the subject has the t(4;14) translocation, the subject is predicted to benefit from treatment with an NSD2 inhibitor.

[0135] In some embodiments, there is provided a method of predicting whether a subject having multiple myeloma will benefit from treatment with an NSD2 inhibitor, comprising: a) obtaining a sample from a subject; b) determining whether the subject has a 4:14 chromosomal translocation (t(4;14)); c) predicting that the subject will benefit from treatment with an NSD2 inhibitor if the subject has a t(4;14) translocation Provided herein is a method comprising:

[0136] Also provided herein are methods for treating patients who have been previously treated for multiple myeloma but are non-responsive to standard therapy, as well as patients who have not been previously treated.Further included are methods for treating patients who have undergone surgery to treat multiple myeloma, as well as patients who have not been previously treated.Also provided herein are methods for treating patients who have undergone transplant therapy, as well as patients who have not been previously treated.

[0137] The methods provided herein include treatment of relapsed, refractory or resistant multiple myeloma. The methods provided herein include prevention of relapsed, refractory or resistant multiple myeloma. The methods provided herein include management of relapsed, refractory or resistant multiple myeloma. In some such embodiments, the myeloma is primary, secondary, tertiary, quaternary or quinary relapsed multiple myeloma. In some embodiments, the methods provided herein reduce, maintain or eliminate minimal residual disease (MRD). In some embodiments, the methods provided herein increase the proportion and / or persistence of MRD negativity in multiple myeloma patients, comprising administering an inhibitor of NSD2 to a subject with multiple myeloma. In some embodiments, the methods provided herein include treating, preventing or managing various types of multiple myeloma, such as monoclonal gammopathy of undetermined significance (MGUS), low-risk, intermediate-risk and high-risk multiple myeloma, newly diagnosed multiple myeloma (including low-risk, intermediate-risk and high-risk newly diagnosed multiple myeloma), transplant eligible and transplant ineligible multiple myeloma, smoldering (low-grade) multiple myeloma (including low-risk, intermediate-risk and high-risk smoldering multiple myeloma), active multiple myeloma, solitary plasmacytoma, extramedullary plasmacytoma, plasma cell leukemia, central nervous system multiple myeloma, light chain myeloma, non-secretory myeloma, immunoglobulin D myeloma and immunoglobulin E myeloma, by administering an inhibitor of NSD2 to a subject with multiple myeloma.

[0138] In another embodiment, the methods provided herein involve the detection of a cyclin D translocation other than, or in addition to, a t(4;14); t(11;14)(q13;q32); t(6;14)(p21;32); t(12;14)(p13;q32); or t(6;20);; an MMSET translocation (e.g., t(4;14)(p16;q32); an MAF translocation (e.g., t(14;16)(q32;q3 2);t(20;22);t(16;22)(q11;q13);or t(14;20)(q32;q11));or other chromosomal factors (17p13 or chromosome 13 deletion;del(17 / 17p), non-hyperdiploidy, and gain(1q)) by administering an inhibitor of NSD2. In some embodiments, the multiple myeloma is characterized according to the International Staging System for Multiple Myeloma (ISS). In some embodiments, the multiple myeloma is stage I multiple myeloma characterized by ISS (e.g., serum β2 microglobulin < 3.5 mg / L and serum albumin ≧ 3.5 g / dL). In some embodiments, the multiple myeloma is stage III multiple myeloma characterized by ISS (e.g., serum β2 microglobulin > 5.4 mg / L). In some embodiments, the multiple myeloma is stage II multiple myeloma (eg, not stage I or III) characterized by ISS.

[0139] In some embodiments, the method comprises administering an inhibitor of NSD2. In some embodiments, the method comprises administering an inhibitor of NSD2 as a consolidation therapy. In some embodiments, the method comprises administering an inhibitor of NSD2 as a maintenance therapy.

[0140] In one particular embodiment of the methods described herein, the multiple myeloma is plasma cell leukemia.

[0141] In some embodiments, the multiple myeloma is high-risk multiple myeloma. In some such embodiments, the high-risk multiple myeloma is relapsed or refractory. In some embodiments, the high-risk multiple myeloma is multiple myeloma that relapsed within 12 months of first treatment. In some embodiments, the high-risk multiple myeloma is multiple myeloma that is further characterized by one or more of genetic abnormalities, such as del(17 / 17p) and t(14;16)(q32;q32). In some such embodiments, the high-risk multiple myeloma is relapsed or refractory to one, two or three previous treatments.

[0142] In some embodiments, the multiple myeloma is further characterized by a p53 mutation. In some embodiments, the p53 mutation is a Q331 mutation. In some embodiments, the p53 mutation is a R273H mutation. In some embodiments, the p53 mutation is a K132 mutation. In some embodiments, the p53 mutation is a K132N mutation. In some embodiments, the p53 mutation is a R337 mutation. In some embodiments, the p53 mutation is a R337L mutation. In some embodiments, the p53 mutation is a W146 mutation. In some embodiments, the p53 mutation is a S261 mutation. In some embodiments, the p53 mutation is a S261T mutation. In some embodiments, the p53 mutation is an E286 mutation. In some embodiments, the p53 mutation is an E286K mutation. In some embodiments, the p53 mutation is a R175 mutation. In some embodiments, the p53 mutation is a R175H mutation. In some embodiments, the p53 mutation is an E258 mutation. In some embodiments, the p53 mutation is an E258K mutation. In some embodiments, the p53 mutation is an A161 mutation. In some embodiments, the p53 mutation is an A161T mutation.

[0143] In some embodiments, the multiple myeloma is characterized by a homozygous deletion of p53. In some embodiments, the multiple myeloma is characterized by a homozygous deletion of wild-type p53.

[0144] In some embodiments, the multiple myeloma is characterized by wild-type p53.

[0145] In some embodiments, the multiple myeloma is characterized by the activation of one or more oncogenic drivers. In some embodiments, the one or more oncogenic drivers are selected from the group consisting of C-MAF, MAFB, FGFR3, MMset, cyclin D1, and cyclin D. In some embodiments, the multiple myeloma is characterized by the activation of C-MAF. In some embodiments, the multiple myeloma is characterized by the activation of MAFB. In some embodiments, the multiple myeloma is characterized by the activation of FGFR3 and MMset. In some embodiments, the multiple myeloma is characterized by the activation of C MAF, FGFR3, and MMset. In certain embodiments, the multiple myeloma is characterized by the activation of cyclin D1. In some embodiments, the multiple myeloma is characterized by the activation of MAFB and cyclin D1. In some embodiments, the multiple myeloma is characterized by the activation of cyclin D.

[0146] In some embodiments, the multiple myeloma is characterized by one or more chromosomal translocations other than or in addition to the chromosomal translocation t(4;14). In some embodiments, the chromosomal translocation is t(4;14) and t(14;16). In some embodiments, the chromosomal translocation is t(4;14) and t(14;20). In some embodiments, the chromosomal translocation is t(4;14) and t(11;14). In some embodiments, the chromosomal translocation is t(4;14) and t(6;20). In some embodiments, the chromosomal translocation is t(4;14) and t(20;22). In some embodiments, the chromosomal translocation is t(4;14) and t(6;20) and t(20;22). In some embodiments, the chromosomal translocation is t(4;14) and t(16;22). In some embodiments, the chromosomal translocation is t(4;14) and t(14;16) and t(16;22). In some embodiments, the chromosomal translocation is t(4;14) and t(14;20) and t(11;14).

[0147] In some embodiments, multiple myeloma is characterized by Q331 p53 mutation, by activation of C-MAF, and by chromosomal translocation at t(4;14) and t(14;16). In some embodiments, multiple myeloma is characterized by homozygous deletion of p53, by activation of C-MAF, and by chromosomal translocation at t(4;14) and t(14;16). In some embodiments, multiple myeloma is characterized by K132N p53 mutation, by activation of MAFB, and by chromosomal translocation at t(4;14) and t(14;20). In some embodiments, multiple myeloma is characterized by wild-type p53, by activation of FGFR3 and MMset, and by chromosomal translocation at t(4;14). In some embodiments, the multiple myeloma is characterized by wild-type p53, activation of C-MAF, and chromosomal translocations at t(4;14) and t(14;16). In some embodiments, the multiple myeloma is characterized by homozygous deletion of p53, activation of FGFR3, MMset, and C-MAF, and chromosomal translocations at t(4;14) and t(14;16). In some embodiments, the multiple myeloma is characterized by homozygous deletion of p53, activation of cyclin D1, and chromosomal translocations at t(4;14) and t(11;14). In some embodiments, the multiple myeloma is characterized by R337L p53 mutation, activation of cyclin D1, and chromosomal translocations at t(4;14) and t(11;14). In some embodiments, the multiple myeloma is characterized by W146 p53 mutation, by activation of FGFR3 and MMset, and by chromosomal translocations at t(4;14) and t(4;14).In some embodiments, the multiple myeloma is characterized by S261T p53 mutation, by activation of MAFB, and by chromosomal translocations at t(4;14) and t(6;20) and t(20;22).In some embodiments, multiple myeloma is characterized by E286K p53 mutation, by activation of FGFR3 and MMset, and by chromosomal translocation at t(4;14). In some embodiments, multiple myeloma is characterized by R175H p53 mutation, by activation of FGFR3 and MMset, and by chromosomal translocation at t(4;14). In some embodiments, multiple myeloma is characterized by E258K p53 mutation, by activation of C-MAF, and by chromosomal translocation at t(4;14) and t(14;16) and t(16;22). In some embodiments, multiple myeloma is characterized by wild-type p53, by activation of MAFB and cyclin D1, and by chromosomal translocation at t(4;14) and t(14;20) and t(11;14). In some embodiments, the multiple myeloma is characterized by an A161T p53 mutation, by activation of cyclin D, and by chromosomal translocations at t(4;14) and t(11;14).

[0148] In some embodiments of the methods described herein, the multiple myeloma is transplant eligible newly diagnosed multiple myeloma. In other embodiments, the multiple myeloma is transplant ineligible newly diagnosed multiple myeloma.

[0149] In yet other embodiments, the multiple myeloma is characterized by early progression (e.g., less than 12 months) after initial treatment. In yet other embodiments, the multiple myeloma is characterized by early progression (e.g., less than 12 months) after autologous stem cell transplant. In some embodiments, the multiple myeloma is refractory to lenalidomide. In some embodiments, the multiple myeloma is refractory to pomalidomide. In some such embodiments, the multiple myeloma is predicted to be refractory to pomalidomide (e.g., by molecular characterization). In some embodiments, the multiple myeloma is relapsed or refractory to three or more therapies and has been exposed to a proteasome inhibitor (e.g., bortezomib, carfilzomib, ixazomib, oprozomib, or marizomib) and an immunomodulatory compound (e.g., thalidomide, lenalidomide, pomalidomide, iverdimide, or avadomide), or is dual refractory to a proteasome inhibitor and an immunomodulatory compound. In yet other embodiments, the multiple myeloma is relapsed or refractory to three or more prior therapies including, for example, a CD38 monoclonal antibody (CD38mAb, e.g., daratumumab or isatuximab), a proteasome inhibitor (e.g., bortezomib, carfilzomib, ixazomib, or marizomib), and an immunomodulatory compound (e.g., thalidomide, lenalidomide, pomalidomide, iveldmide, or avadomide), or is dual refractory to a proteasome inhibitor or immunomodulatory compound and a CD38mAb. In yet other embodiments, the multiple myeloma is triple refractory, for example, the multiple myeloma is refractory to a proteasome inhibitor (e.g., bortezomib, carfilzomib, ixazomib, oprozomib, or marizomib), an immunomodulatory compound (e.g., thalidomide, lenalidomide, pomalidomide, iverdomib, or avadomide), and one other active agent described herein.

[0150] In certain embodiments, provided herein is a method of treating, preventing, and / or managing multiple myeloma, including relapsed / refractory multiple myeloma, in a subject with renal dysfunction or symptoms thereof, comprising administering an inhibitor of NSD2 to a subject with relapsed / refractory multiple myeloma with renal dysfunction.

[0151] In certain embodiments, the present disclosure provides a method for treating, preventing, and / or managing multiple myeloma, including relapsed / refractory multiple myeloma, in frail subjects, comprising administering an inhibitor of NSD2 to the frail subject with multiple myeloma.In some such embodiments, the frail subject is characterized by being ineligible for induction therapy or being intolerant to dexamethasone treatment.In some such embodiments, the frail subject is elderly, for example, older than 65 years old.

[0152] In certain embodiments, provided herein is a method of treating, preventing or managing multiple myeloma, comprising administering to a subject an inhibitor of NSD2, wherein the multiple myeloma is quaternary relapsed / refractory multiple myeloma.

[0153] In certain embodiments, provided herein is a method of treating, preventing or managing multiple myeloma, comprising administering to a subject an inhibitor of NSD2 as induction therapy, wherein the multiple myeloma is newly diagnosed, transplant-eligible multiple myeloma.

[0154] In certain embodiments, provided herein is a method of treating, preventing or managing multiple myeloma, comprising administering to a subject an NSD2 inhibitor as a maintenance therapy following other treatment or transplant, wherein the multiple myeloma is newly diagnosed transplant-eligible multiple myeloma prior to other treatment or transplant.

[0155] In certain embodiments, the present disclosure provides a method for treating, preventing or managing multiple myeloma, comprising administering to a subject an inhibitor of NSD2 as a maintenance therapy after other treatment or transplantation.In some embodiments, the multiple myeloma is a newly diagnosed transplant-eligible multiple myeloma before other treatment and / or transplantation.In some embodiments, the other treatment before transplantation is chemotherapy or treatment with an inhibitor of NSD2.

[0156] In certain embodiments, provided herein is a method of treating, preventing or managing multiple myeloma, comprising administering to a subject an inhibitor of NSD2, wherein the multiple myeloma is high-risk multiple myeloma that is relapsed or refractory to one, two or three prior treatments.

[0157] In certain embodiments, provided herein is a method of treating, preventing or managing multiple myeloma, comprising administering to a subject an inhibitor of NSD2, wherein the multiple myeloma is newly diagnosed transplant-ineligible multiple myeloma.

[0158] In certain embodiments, the subject to be treated with one of the methods provided herein has not been treated with multiple myeloma therapy before administering the inhibitor of NSD2.In certain embodiments, the subject to be treated with one of the methods provided herein has been treated with multiple myeloma therapy before administering the inhibitor of NSD2.In certain embodiments, the subject to be treated with one of the methods provided herein has developed drug resistance to anti-multiple myeloma therapy.In some such embodiments, the subject has developed resistance to one, two, or three anti-multiple myeloma therapy, and the therapy is selected from CD38 monoclonal antibody (CD38 mAb, for example, daratumumab or isatuximab), proteasome inhibitor (for example, bortezomib, carfilzomib, ixazomib, or marizomib), and immunomodulatory compound (for example, thalidomide, lenalidomide, pomalidomide, iveldomide, or avadomide).

[0159] The methods provided herein encompass treating subjects regardless of the patient's age. In some embodiments, the subject is at least 18 years old or older. In other embodiments, the subject is older than 18, 25, 35, 40, 45, 50, 55, 60, 65, or 70 years old. In other embodiments, the subject is younger than 65 years old. In other embodiments, the subject is older than 65 years old. In some embodiments, the subject is an elderly multiple myeloma subject, such as a subject older than 65 years old. In some embodiments, the subject is an elderly multiple myeloma subject, such as a subject older than 75 years old.

[0160] E. Combination Therapy with At Least One Second Therapeutic Agent In some embodiments, the methods provided herein (use of an inhibitor of NSD2) further comprise administering to the subject at least one second therapeutic agent, also referred to herein as an "additional agent" or "additional active agent."

[0161] In some embodiments, the at least one second therapeutic agent is selected from a chemotherapeutic agent, a steroid, an immunomodulatory agent, a proteasome inhibitor, a histone deacetylase inhibitor, an anti-CD38 antibody, and an anti-SLAMF7 antibody, an antibody drug conjugate, and a nuclear export inhibitor.

[0162] In some embodiments, the at least one second therapeutic agent is selected from lenalidomide, thalidomide, pomalidomide, bortezomib, carfilzomib, ixazomib, panobinostat, melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, and bendamustine, dexamethasone, prednisone, daratumumab, isatuximab, elotuzumab, belantamab mafodotin-blmf, selinexor, pamidronate, zoledronic acid, and denosumab.

[0163] In some embodiments, the at least one second therapeutic agent is lenalidomide. In some embodiments, the at least one second therapeutic agent is iverdomi. In some embodiments, the at least one second therapeutic agent is (S)-4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)methyl)benzyl)piperazin-1-yl)-3-fluorobenzonitrile. In some embodiments, the at least one second therapeutic agent is (i) lenalidomide, pomalidomide, or thalidomide; and (ii) dexamethasone. In some embodiments, the at least one second therapeutic agent is (i) carfilzomib, ixazomib, or bortezomib; (ii) lenalidomide; and (iii) dexamethasone. In some embodiments, the at least one second therapeutic agent is (i) bortezomib or carfilzomib; (ii) cyclophosphamide; and (iii) dexamethasone. In some embodiments, the at least one second therapeutic agent is (i) elotuzumab or daratumumab; (ii) lenalidomide; and (iii) dexamethasone. In some embodiments, the at least one second therapeutic agent is bortezomib, liposomal doxorubicin, and dexamethasone. In some embodiments, the at least one second therapeutic agent is panobinostat, bortezomib, and dexamethasone. In some embodiments, the at least one second therapeutic agent is elotuzumab, bortezomib, and dexamethasone. In some embodiments, the at least one second therapeutic agent is melphalan and prednisone, with or without thalidomide or bortezomib. In some embodiments, at least one second therapeutic agent is vincristine, doxorubicin, and dexamethasone.In some embodiments, at least one second therapeutic agent is dexamethasone, cyclophosphamide, etoposide, and cisplatin.In some embodiments, at least one second therapeutic agent is dexamethasone, thalidomide, cisplatin, doxorubicin, cyclophosphamide, and etoposide, with or without bortezomib.

[0164] In some embodiments, the at least one second therapeutic agent is a steroid.

[0165] In some embodiments, the specific amount (dosage) of at least one second therapeutic agent provided herein used in the methods provided herein will be determined by factors such as the specific agent used, the type of multiple myeloma being treated or managed, the severity and stage of the disease, the amount of an inhibitor of NSD2 provided herein, and any optional additional active agents co-administered to the patient.

[0166] In some embodiments, the dosage of at least one second therapeutic agent provided herein used in the methods provided herein is determined based on the commercially available pharmaceutical package insert (e.g., label) approved by the FDA or a similar regulatory agency in a country other than the United States for said active agent. In some embodiments, the dosage of the second therapeutic agent provided herein used in the methods provided herein is the dosage approved by the FDA or a similar regulatory agency in a country other than the United States for said active agent. In some embodiments, the dosage of the second therapeutic agent provided herein used in the methods provided herein is the dosage used in a human clinical trial of said active agent. In some embodiments, the dosage of the second therapeutic agent provided herein used in the methods provided herein is lower than the dosage approved by the FDA or a similar regulatory agency in a country other than the United States for said active agent or the dosage used in a human clinical trial of said active agent, for example, depending on the synergistic effect between the second therapeutic agent and the inhibitor of NSD2 provided herein.

[0167] Combinations of NSD2 inhibitors provided herein can also be combined or used with (e.g., before, during, or after) conventional therapy, including, but not limited to, surgery, biological therapy (e.g., immunotherapy with checkpoint inhibitors), radiation therapy, chemotherapy, stem cell transplantation, cell therapy, or other non-pharmacologic therapy currently used to treat, prevent, or manage cancer (e.g., multiple myeloma). Combinations of NSD2 inhibitors provided herein with conventional therapy can provide unique treatment regimens that are unexpectedly effective in certain patients. Without being limited to a particular theory, it is believed that NSD2 inhibitors provided herein can provide additive or synergistic effects when administered simultaneously with conventional therapy.

[0168] As discussed elsewhere herein, methods are included herein for reducing, treating and / or preventing the harmful or undesirable effects associated with conventional therapy, including but not limited to surgery, chemotherapy, radiation therapy, biological therapy and immunotherapy.The inhibitor of NSD2 provided herein and at least one second therapeutic agent component can be administered to patients before, during or after the occurrence of the harmful effects associated with conventional therapy.In some embodiments, at least one second therapeutic agent is dexamethasone.

[0169] In addition, the inhibitors of NSD2 provided herein can be further combined or used in combination with other therapeutic agents useful for the treatment and / or prevention of multiple myeloma as described herein. In one such embodiment, at least one second therapeutic agent is dexamethasone.

[0170] In some embodiments, provided herein are methods for treating, preventing, or managing multiple myeloma, comprising administering to a patient an inhibitor of NSD2 provided herein, further in combination with one or more additional therapeutic agents, and optionally further in combination with radiation therapy, blood transfusion, or surgery.

[0171] As used herein, the term "in combination" includes the use of more than one therapies (e.g., one or more prophylactic and / or therapeutic agents). However, the use of the term "in combination" does not restrict the order in which therapies (e.g., prophylactic and / or therapeutic agents) are administered to a patient with a disease or disorder. The first therapy (e.g., a prophylactic or therapeutic agent, such as an inhibitor of NSD2 provided herein) may be administered prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks prior to) administration of the second therapy (e.g., at least one second therapeutic agent) to the subject, concurrently with administration of the second therapy (e.g., at least one second therapeutic agent) to the subject, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) administration of the second therapy (e.g., at least one second therapeutic agent) to the subject. The first and second treatments can be administered independently prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before) the administration of the third treatment (e.g., an additional prophylactic or therapeutic agent) to the subject, simultaneously with (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of the third treatment (e.g., an additional prophylactic or therapeutic agent) to the subject. Four-drug combinations are also contemplated herein, and five-drug combinations are also contemplated. In some embodiments, the second treatment is dexamethasone.

[0172] The administration of the inhibitor of NSD2 provided herein and one or more second therapeutic agents to a subject can be performed simultaneously or sequentially by the same or different routes of administration.The suitability of the particular route of administration used for a particular therapeutic agent will depend on the active agent itself (e.g., whether it can be administered orally without being decomposed before entering the bloodstream) and the cancer to be treated.

[0173] The route of administration of the inhibitor of NSD2 provided herein is independent of the additional treatment. In some embodiments, the inhibitor of NSD2 provided herein is administered orally. In another embodiment, the inhibitor of NSD2 provided herein is administered intravenously. Thus, according to these embodiments, the inhibitor of NSD2 provided herein is administered orally or intravenously, and the additional treatment can be administered orally, parenterally, intraperitoneally, intravenously, intraarterially, transdermally, sublingually, intramuscularly, rectally, buccally, intranasally, via liposomes, via inhalation, intravaginally, intraocularly, via local delivery by catheter or stent, subcutaneously, intraadiposally, intraarticularly, intrathecally, or in sustained release dosage form. In some embodiments, the inhibitor of NSD2 provided herein and the additional treatment are administered by the same mode of administration, orally, or by IV. In another embodiment, the inhibitor of NSD2 provided herein is administered by one mode of administration, e.g., by IV, while the additional agent (an anti-multiple myeloma agent) is administered by another mode of administration, e.g., orally.

[0174] In some embodiments, the at least one second therapeutic agent is administered intravenously or subcutaneously, and in an amount of about 1 to about 1000 mg, about 5 to about 500 mg, about 10 to about 350 mg, or about 50 to about 200 mg, once or twice daily. The specific amount of the additional active agent will depend on the particular agent used, the type of multiple myeloma being treated and / or managed, the severity and stage of the disease, the amount of the inhibitor of NSD2 provided herein, and any optional additional active agents co-administered to the subject.

[0175] One or more additional active ingredients or agents may be used in the methods and compositions provided herein with the inhibitors of NSD2 provided herein. The additional active agent may be a large molecule (e.g., a protein) or a small molecule (e.g., a synthetic inorganic, organometallic, or organic molecule) or a cell therapy (e.g., CAR cells).

[0176] Examples of additional active agents that can be used in the methods and compositions described herein include melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, bendamustine, obinutuzumab, proteasome inhibitors (e.g., bortezomib, carfilzomib, ixazomib, oprozomib, or marizomib), deacetylase inhibitors such as histone deacetylase inhibitors (as described herein, e.g., panobinostat, ACY241), BET inhibitors (e.g., GSK525762A, OTX015, BMS -986158, TEN-010, CPI-0610, INCB54329, BAY1238097, FT-1101, ABBV-075, BI894999, GS-5829, GSK1210151A (I-BET-151), CPI-203, RVX-208, XD46, MS436, PFI-1, RVX2135, ZEN3365, XD14, ARV-771, MZ-1, PLX5117, 4-[2-(cyclopropylmethoxy)-5-(methanesulfonyl)phenyl]-2-methylisoquinolin-1(2H)-one, EP113 13 and EP11336), BCL2 inhibitors (e.g., venetoclax or navitoclax), MCL-1 inhibitors (e.g., AZD5991, AMG176, MIK665, S64315, or S63845), LSD-1 inhibitors (e.g., ORY-1001, ORY-2001, INCB-59872, IMG-7289, TAK-418, GSK-2879552, 4-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydropyrimidine -4-yl]-2-fluoro-benzonitrile or salt thereof), a corticosteroid (e.g., prednisone), dexamethasone; an antibody (e.g., a CS1 antibody such as elotuzumab, a CD38 antibody such as daratumumab or isatuximab); or a BCMA antibody or antibody conjugate such as GSK2857916 or BI836909), a checkpoint inhibitor (as described herein), or a CAR cell (as described herein); or an anti-SLAMF7 antibody (as described herein); or an antibody drug conjugate (as described herein);or one or more of the nuclear export inhibitors (described herein);

[0177] In some embodiments, the additional active agent used in conjunction with the inhibitors of NSD2 provided herein in the methods and compositions described herein is dexamethasone.

[0178] In some embodiments, dexamethasone is administered at a dose of 4 mg on days 1 and 8 of a 21-day cycle. In some other embodiments, dexamethasone is administered at a dose of 4 mg on days 1, 4, 8, and 11 of a 21-day cycle. In some embodiments, dexamethasone is administered at a dose of 4 mg on days 1, 8, and 15 of a 28-day cycle. In some other embodiments, dexamethasone is administered at a dose of 4 mg on days 1, 4, 8, 11, 15, and 18 of a 28-day cycle. In some embodiments, dexamethasone is administered at a dose of 4 mg on days 1, 8, 15, and 22 of a 28-day cycle. In one such embodiment, dexamethasone is administered at a dose of 4 mg on days 1, 10, 15, and 22 of cycle 1. In some embodiments, dexamethasone is administered at a dose of 4 mg on days 1, 3, 15, and 17 of a 28 day cycle. In one such embodiment, dexamethasone is administered at a dose of 4 mg on days 1, 3, 14, and 17 of cycle 1.

[0179] In some other embodiments, dexamethasone is administered at a dose of 8 mg on days 1 and 8 of a 21-day cycle. In some other embodiments, dexamethasone is administered at a dose of 8 mg on days 1, 4, 8, and 11 of a 21-day cycle. In some embodiments, dexamethasone is administered at a dose of 8 mg on days 1, 8, and 15 of a 28-day cycle. In some other embodiments, dexamethasone is administered at a dose of 8 mg on days 1, 4, 8, 11, 15, and 18 of a 28-day cycle. In some embodiments, dexamethasone is administered at a dose of 8 mg on days 1, 8, 15, and 22 of a 28-day cycle. In one such embodiment, dexamethasone is administered at a dose of 8 mg on days 1, 10, 15, and 22 of cycle 1. In some embodiments, dexamethasone is administered at a dose of 8 mg on days 1, 3, 15, and 17 of a 28 day cycle. In one such embodiment, dexamethasone is administered at a dose of 8 mg on days 1, 3, 14, and 17 of cycle 1.

[0180] In some embodiments, dexamethasone is administered at a dose of 10 mg on days 1 and 8 of a 21-day cycle. In some other embodiments, dexamethasone is administered at a dose of 10 mg on days 1, 4, 8, and 11 of a 21-day cycle. In some embodiments, dexamethasone is administered at a dose of 10 mg on days 1, 8, and 15 of a 28-day cycle. In some other embodiments, dexamethasone is administered at a dose of 10 mg on days 1, 4, 8, 11, 15, and 18 of a 28-day cycle. In some embodiments, dexamethasone is administered at a dose of 10 mg on days 1, 8, 15, and 22 of a 28-day cycle. In some embodiments, dexamethasone is administered at a dose of 10 mg on days 1, 10, 15, and 22 of cycle 1. In some embodiments, dexamethasone is administered at a dose of 10 mg on days 1, 3, 15, and 17 of a 28 day cycle. In some embodiments, dexamethasone is administered at a dose of 10 mg on days 1, 3, 14, and 17 of cycle 1.

[0181] In some embodiments, dexamethasone is administered at a dose of 20 mg on days 1 and 8 of a 21-day cycle. In some other embodiments, dexamethasone is administered at a dose of 20 mg on days 1, 4, 8, and 11 of a 21-day cycle. In some embodiments, dexamethasone is administered at a dose of 20 mg on days 1, 8, and 15 of a 28-day cycle. In some other embodiments, dexamethasone is administered at a dose of 20 mg on days 1, 4, 8, 11, 15, and 18 of a 28-day cycle. In some embodiments, dexamethasone is administered at a dose of 20 mg on days 1, 8, 15, and 22 of a 28-day cycle. In some embodiments, dexamethasone is administered at a dose of 20 mg on days 1, 10, 15, and 22 of cycle 1. In some embodiments, dexamethasone is administered at a dose of 20 mg on days 1, 3, 15, and 17 of a 28 day cycle. In some embodiments, dexamethasone is administered at a dose of 20 mg on days 1, 3, 14, and 17 of cycle 1.

[0182] In some embodiments, dexamethasone is administered at a dose of 40 mg on days 1 and 8 of a 21-day cycle. In some other embodiments, dexamethasone is administered at a dose of 40 mg on days 1, 4, 8, and 11 of a 21-day cycle. In some embodiments, dexamethasone is administered at a dose of 40 mg on days 1, 8, and 15 of a 28-day cycle. In one such embodiment, dexamethasone is administered at a dose of 40 mg on days 1, 10, 15, and 22 of cycle 1. In some other embodiments, dexamethasone is administered at a dose of 40 mg on days 1, 4, 8, 11, 15, and 18 of a 28-day cycle. In other such embodiments, dexamethasone is administered at a dose of 40 mg on days 1, 8, 15, and 22 of a 28-day cycle. In other such embodiments, dexamethasone is administered at a dose of 40 mg on days 1, 3, 15, and 17 of a 28 day cycle. In some embodiments, dexamethasone is administered at a dose of 40 mg on days 1, 3, 14, and 17 of cycle 1.

[0183] In another embodiment, the additional active agent used in the methods and compositions described herein with the inhibitor of NSD2 provided herein is bortezomib.In yet another embodiment, the additional active agent used in the methods and compositions described herein with the inhibitor of NSD2 provided herein is daratumumab.In some such embodiments, the method further comprises administering dexamethasone.In some embodiments, the method comprises administering the NSD2 inhibitor provided herein with a proteasome inhibitor described herein, a CD38 inhibitor described herein, and a corticosteroid described herein.

[0184] In certain embodiments, the inhibitor of NSD2 provided herein is administered in combination with a checkpoint inhibitor.In some embodiments, one checkpoint inhibitor is used in combination with the inhibitor of NSD2 provided herein in connection with the methods provided herein.In another embodiment, two checkpoint inhibitors are used in combination with the inhibitor of NSD2 provided herein in connection with the methods provided herein.In yet another embodiment, three or more checkpoint inhibitors are used in combination with the inhibitor of NSD2 provided herein in connection with the methods provided herein.

[0185] As used herein, the term "immune checkpoint inhibitor" or "checkpoint inhibitor" refers to a molecule that completely or partially reduces, inhibits, interferes with or modulates one or more checkpoint proteins. Without being limited to a particular theory, checkpoint proteins control the activation or function of T cells. Many checkpoint proteins are known, such as CTLA-4 and its ligands CD80 and CD86; and PD-1 and its ligands PD-L1 and PD-L2 (Pardoll, Nature Reviews Cancer, 2012, 12, 252-264). These proteins appear to be involved in costimulatory or inhibitory interactions of T cell responses. Immune checkpoint proteins appear to control and maintain self-tolerance as well as the duration and breadth of physiological immune responses. Immune checkpoint inhibitors include or are derived from antibodies.

[0186] In some embodiments, the checkpoint inhibitor is a CTLA-4 inhibitor. In some embodiments, the CTLA-4 inhibitor is an anti-CTLA-4 antibody. Examples of anti-CTLA-4 antibodies include, but are not limited to, those described in U.S. Patent Nos. 5,811,097; 5,811,097; 5,855,887; 6,051,227; 6,207,157; ​​6,682,736; 6,984,720; and 7,605,238, all of which are incorporated herein in their entirety. In some embodiments, the anti-CTLA-4 antibody is tremelimumab (also known as ticilimumab or CP-675,206). In another embodiment, the anti-CTLA-4 antibody is ipilimumab (also known as MDX-010 or MDX-101). Ipilimumab is a fully human monoclonal IgG antibody that binds to CTLA-4. Ipilimumab is commercially available under the trade name Yervoy™.

[0187] In some embodiments, the checkpoint inhibitor is a PD-1 / PD-L1 inhibitor. Examples of PD-1 / PD-L1 inhibitors include, but are not limited to, those described in U.S. Patent Nos. 7,488,802; 7,943,743; 8,008,449; 8,168,757; 8,217,149, and PCT Patent Application Publication Nos. WO2003042402, WO2008156712, WO2010089411, WO2010036959, WO2011066342, WO2011159877, WO2011082400, and WO2011161699, all of which are incorporated herein in their entirety.

[0188] In some embodiments, the checkpoint inhibitor is a PD-1 inhibitor. In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 antibody is BGB-A317, nivolumab (also known as ONO-4538, BMS-936558, or MDX1106) or pembrolizumab (also known as MK-3475, SCH900475, or ramobrolizumab). In some embodiments, the anti-PD-1 antibody is nivolumab. Nivolumab is a human IgG4 anti-PD-1 monoclonal antibody and is commercially available under the trade name Opdivo™. In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 antibody and is commercially available under the trade name Keytruda™. In yet another embodiment, the anti-PD-1 antibody is the humanized antibody CT-011. Administration of CT-011 alone has not shown a response in the treatment of acute myeloid leukemia (AML) at relapse. In yet another embodiment, the anti-PD-1 antibody is the fusion protein AMP-224. In another embodiment, the PD-1 antibody is BGB-A317. BGB-A317 is a monoclonal antibody that has been specifically engineered for its ability to bind to Fc-gamma receptor I and has a unique binding signature to PD-1 with high affinity and excellent target specificity.

[0189] In some embodiments, the checkpoint inhibitor is a PD-L1 inhibitor. In some embodiments, the PD-L1 inhibitor is an anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody is MEDI4736 (durvalumab). In another embodiment, the anti-PD-L1 antibody is BMS-936559 (also known as MDX-1105-01). In yet another embodiment, the PD-L1 inhibitor is atezolizumab (also known as MPDL3280A, and Tecentriq®).

[0190] In some embodiments, the checkpoint inhibitor is a PD-L2 inhibitor. In some embodiments, the PD-L2 inhibitor is an anti-PD-L2 antibody. In some embodiments, the anti-PD-L2 antibody is rHIgM12B7A.

[0191] In some embodiments, the checkpoint inhibitor is lymphocyte activation gene 3 (LAG-3) inhibitor. In some embodiments, the LAG-3 inhibitor is soluble Ig fusion protein IMP321 (Brignone et al., J. Immunol., 2007, 179, 4202-4211). In another embodiment, the LAG-3 inhibitor is BMS-986016.

[0192] In some embodiments, the checkpoint inhibitor is a B7 inhibitor. In some embodiments, the B7 inhibitor is a B7-H3 inhibitor or a B7-H4 inhibitor. In some embodiments, the B7-H3 inhibitor is an anti-B7-H3 antibody MGA271 (Loo et al., Clin. Cancer Res., 2012, 3834).

[0193] In some embodiments, the checkpoint inhibitor is a TIM3 (T-cell immunoglobulin domain and mucin domain 3) inhibitor (Fourcade et al., J. Exp. Med., 2010, 207, 2175-86; Sakuishi et al., J. Exp. Med., 2010, 207, 2187-94).

[0194] In some embodiments, the checkpoint inhibitor is an OX40 (CD134) agonist. In some embodiments, the checkpoint inhibitor is an anti-OX40 antibody. In some embodiments, the anti-OX40 antibody is anti-OX-40. In another embodiment, the anti-OX40 antibody is MEDI6469.

[0195] In some embodiments, the checkpoint inhibitor is a GITR agonist. In some embodiments, the checkpoint inhibitor is an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518.

[0196] In some embodiments, the checkpoint inhibitor is a CD137 agonist. In some embodiments, the checkpoint inhibitor is an anti-CD137 antibody. In some embodiments, the anti-CD137 antibody is urelumab. In another embodiment, the anti-CD137 antibody is PF-05082566.

[0197] In some embodiments, the checkpoint inhibitor is a CD40 agonist. In some embodiments, the checkpoint inhibitor is an anti-CD40 antibody. In some embodiments, the anti-CD40 antibody is CF870,893.

[0198] In some embodiments, the checkpoint inhibitor is recombinant human interleukin-15 (rhIL-15).

[0199] In some embodiments, the checkpoint inhibitor is an IDO inhibitor. In some embodiments, the IDO inhibitor is INCB024360. In another embodiment, the IDO inhibitor is indoximod.

[0200] In certain embodiments, the combination therapy provided herein comprises two or more of the checkpoint inhibitors described herein (including the same or different types of checkpoint inhibitors).Furthermore, the combination therapy described herein can be used in combination with one or more second therapeutic agents described herein, if appropriate for the treatment of the diseases described herein and art-recognized diseases.

[0201] In certain embodiments, the inhibitors of NSD2 provided herein may be used in combination with one or more immune cells (e.g., modified immune cells) expressing one or more chimeric antigen receptors (CARs) on their surface. In general, a CAR comprises an extracellular domain from a first protein (e.g., an antigen-binding protein), a transmembrane domain, and an intracellular signaling domain. In certain embodiments, when the extracellular domain binds to a target protein, such as a tumor-associated antigen (TAA) or a tumor-specific antigen (TSA), a signal is generated through the intracellular signaling domain that activates the immune cell, for example, to target and kill cells expressing the target protein.

[0202] Extracellular domain: The extracellular domain of the CAR binds to an antigen of interest. In certain embodiments, the extracellular domain of the CAR comprises a receptor, or a portion of a receptor, that binds to the antigen. In certain embodiments, the extracellular domain comprises or is an antibody or an antigen-binding portion thereof. In certain embodiments, the extracellular domain comprises or is a single-chain Fv (scFv) domain. A single-chain Fv domain may comprise a VL linked to a VH by a flexible linker, for example, when the VL and VH are derived from an antibody that binds to the antigen.

[0203] In certain embodiments, the antigen recognized by the extracellular domain of the polypeptide described herein is a tumor-associated antigen (TAA) or tumor-specific antigen (TSA). In various specific embodiments, the tumor-associated or tumor-specific antigen is, but is not limited to, Her2, prostate stem cell antigen (PSCA), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen-125 (CA-125), CA19-9, calretinin, MUC-1, B-cell maturation antigen (BCMA), epithelial membrane protein (EMA), epithelial tumor antigen (ETA), tyrosinase, melanoma-24 associated antigen (MAGE), CD19, CD22, CD27, CD30, CD34, CD45, CD70, CD99, CD117, EGFRvIII (epidermal growth factor variant III), mesothelin, PAP (prostatic acid phosphatase), prostein, TARP (T-cell receptor gamma alternative reading frame protein), or other antigens. cytokeratin, Trp-p8, STEAPI (six transmembrane epithelial antigen of prostate 1), chromogranin, cytokeratin, desmin, glial fibrillary acidic protein (GFAP), macroscopic cystic disease fluid protein (GCDFP-15), HMB-45 antigen, protein melan-A (melanoma antigen recognized by T lymphocytes; MART-I), myo-D1, muscle specific actin (MSA), neurofilament, neuron specific enolase (NSE), placental alkaline phosphatase, synaptophysis, thyroglobulin, thyroid transcription factor-1, dimeric form of pyruvate kinase isoenzyme type M2 (tumor M2-PK), abnormal ras protein, or abnormal p53 protein. In certain other embodiments, the TAA or TSA recognized by the extracellular domain of the CAR is integrin αvβ (CD61), galactin, or Ral-B.

[0204] In certain embodiments, the TAA or TSA recognized by the extracellular domain of the CAR is a cancer / testis (CT) antigen, such as BAGE, CAGE, CTAGE, FATE, GAGE, HCA661, HOM-TES-85, MAGEA, MAGEB, MAGEC, NA88, NY-ES0-1, NY-SAR-35, OY-TES-1, SPANXBI, SPA17, SSX, SYCPI, or TPTE.

[0205] In certain other embodiments, the TAA or TSA recognized by the extracellular domain of the CAR is a carbohydrate or ganglioside, such as fuc-GM1, GM2 (oncofetal antigen-immunogenic-1; OFA-I-1); GD2 (OFA-I-2), GM3, GD3, etc.

[0206] In certain other embodiments, the TAA or TSA recognized by the extracellular domain of the CAR is alpha-actinin-4, Bage-1, BCR-ABL, Bcr-Ab1 fusion protein, beta-catenin, CA125, CA15-3 (CA27.29\BCAA), CA195, CA242, CA-50, CAM43, Casp-8, cdc27, cdk4, cdkn2a, CEA, coa-1, dek-can fusion protein, EBNA, EF2, Epstein-Barr virus antigen, ETV6-AML1 fusion protein, or a combination thereof. protein, HLA-A2, HLA-All, hsp70-2, KIAA0205, Mart2, Mum-1, 2 and 3, neo-PAP, myosin class I, OS-9, pml-RARα fusion protein, PTPRK, K-ras, N-ras, triosephosphate isomerase, Gage3,4,5,6,7, GnTV, Herv-K-mel, Lage-1, NA-88, NY-Eso-1 / Lage-2, SP17, SSX-2, TRP2-Int2, gp100 (Pmel17), tyrosinase, TRP-1, TR P-2, MAGE-1, MAGE-3, RAGE, GAGE-1, GAGE-2, p15(58), RAGE, SCP-1, Hom / Mel-40, PRAME, p53, HRas, HER-2 / neu, E2A-PRL, H4-RET, IGH-IGK, MY L-RAR, human papillomavirus (HPV) antigens E6 and E7, TSP-180, MAGE-4, MAGE-5, MAGE-6, p185erbB2, p180erbB-3, c-met, nm-23H1, PSA, TAG-72-4, CA19-9, CA 72-4, CAM17.1, NuMa, K-ras, 13-catenin, Mum-1, p16, TAGE, PSMA, CT7, telomerase, 43-9F, 5T4, 791Tgp72, 13HCG, BCA225, BTAA, CD68\KP1, C0-029, FGF-5, G250, Ga733 (EpCAM), HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB\70K, NY-C0-1, RCAS1, SDCCAG16, TA-90, TAAL6, TAG72, TLP, or TPS.

[0207] In various specific embodiments, the tumor-associated or tumor-specific antigen is an AML-associated tumor antigen, as described in S. Anguille et al, Leukemia (2012), 26, 2186-2196.

[0208] Other tumor-associated and tumor-specific antigens are known in the art.

[0209] Receptors, antibodies, and scFvs that bind to TSAs and TAAs and are useful for constructing chimeric antigen receptors are known in the art, as are the nucleotide sequences that encode them.

[0210] In certain embodiments, the antigen recognized by the extracellular domain of the chimeric antigen receptor is not generally considered to be a TSA or a TAA, but is nevertheless an antigen associated with tumor cells or tumor-induced damage.In certain embodiments, for example, the antigen is, for example, a growth factor, cytokine, or interleukin, such as an angiogenesis or vasculogenesis-associated growth factor, cytokine, or interleukin.Such growth factors, cytokines, or interleukins may include, for example, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), or interleukin-8 (IL-8).Tumors can also create a local hypoxic environment for tumors.Therefore, in other certain embodiments, the antigen is a hypoxia-associated factor, for example, HIF-1α, HIF-1β, HIF-2α, HIF-2β, HIF-3α, or HIF-3β. Tumors can also cause localized damage to normal tissues, resulting in the release of molecules known as damage-associated molecular pattern molecules (DAMPs; also known as alarmins). In certain other embodiments, the antigen is thus a DAMP, such as heat shock protein, chromatin-associated protein high mobility group box 1 (HMGB1), S100A8 (MRP8, calgranulin A), S100A9 (MRP14, calgranulin B), serum amyloid A (SAA), or it can be deoxyribonucleic acid, adenosine triphosphate, uric acid, or heparin sulfate.

[0211] Transmembrane domain: In certain embodiments, the extracellular domain of the CAR is linked to the transmembrane domain of the polypeptide by a linker, spacer, or hinge polypeptide sequence, such as a sequence derived from CD28 or a sequence derived from CTLA4. The transmembrane domain can be obtained or derived from the transmembrane domain of any transmembrane protein, and can include all or part of such a transmembrane domain. In certain embodiments, the transmembrane domain can be obtained or derived from, for example, CD8, CD16, cytokine receptors, and interleukin receptors, or growth factor receptors, etc.

[0212] Intracellular signaling domain: In certain embodiments, the intracellular domain of the CAR is expressed on the surface of a T cell and induces activation and / or proliferation of said T cell. It is or comprises an intracellular domain or motif of a protein. Such a domain or motif can transmit a primary antigen binding signal that is necessary for activation of T lymphocytes in response to binding of an antigen to the extracellular portion of the CAR. Typically, this domain or motif comprises or is an ITAM (immunoreceptor tyrosine-based activation motif). Suitable ITAM-containing polypeptides for CARs include, for example, the zeta CD3 chain (CD3ζ) or an ITAM-containing portion thereof. In certain embodiments, the intracellular domain is a CD3ζ intracellular signaling domain. In other specific embodiments, the intracellular domain is derived from a lymphocyte receptor chain, a TCR / CD3 complex protein, an Fe receptor subunit, or an IL-2 receptor subunit. In certain embodiments, the CAR further comprises one or more co-stimulatory domains or motifs, for example, as part of the intracellular domain of the polypeptide. The one or more costimulatory domains or motifs can be or include one or more of a costimulatory CD27 polypeptide sequence, a costimulatory CD28 polypeptide sequence, a costimulatory OX40 (CD134) polypeptide sequence, a costimulatory 4-1BB (CD137) polypeptide sequence, or a costimulatory inducible T cell costimulatory (ICOS) polypeptide sequence, or other costimulatory domains or motifs, or any combination thereof.

[0213] CAR may also include a T cell survival motif. The T cell survival motif may be any polypeptide sequence or motif that promotes the survival of T lymphocytes after stimulation with an antigen. In certain embodiments, the T cell survival motif is or is derived from CD3, CD28, the intracellular signaling domain of IL-7 receptor (IL-7R), the intracellular signaling domain of IL-12 receptor, the intracellular signaling domain of IL-15 receptor, the intracellular signaling domain of IL-21 receptor, or the intracellular signaling domain of transforming growth factor beta (TGFβ) receptor.

[0214] The modified immune cells expressing a CAR can be, for example, T lymphocytes (T cells, e.g., CD4+ T cells or CD8+ T cells), cytotoxic lymphocytes (CTLs), or natural killer (NK) cells. The T lymphocytes used in the compositions and methods provided herein can be naive T lymphocytes or MHC-restricted T lymphocytes. In certain embodiments, the T lymphocytes are tumor-infiltrating lymphocytes (TILs). In certain embodiments, the T lymphocytes are isolated from a tumor biopsy or expanded from T lymphocytes isolated from a tumor biopsy. In certain other embodiments, the T cells are isolated from peripheral blood, umbilical cord blood, or lymph, or expanded from T lymphocytes isolated from peripheral blood, umbilical cord blood, or lymph. The immune cells used to generate the modified immune cells expressing a CAR can be isolated using conventional methods accepted in the art (e.g., apheresis after blood collection and, if appropriate, antibody-mediated cell isolation or sorting).

[0215] The modified immune cells are preferably autologous to the individual to whom the modified immune cells are administered. In certain other embodiments, the modified immune cells are allogeneic to the individual to whom the modified immune cells are administered. When using allogeneic T lymphocytes or NK cells to prepare the modified T lymphocytes, it is preferable to select T lymphocytes or NK cells that will reduce the possibility of graft-versus-host disease (GVHD) in the individual. For example, in certain embodiments, virus-specific T lymphocytes are selected for the preparation of the modified T lymphocytes; such lymphocytes would be considered to have a greatly reduced natural ability to bind to any recipient antigen and thus become activated by the antigen. In certain embodiments, the recipient-mediated rejection of allogeneic T lymphocytes can be reduced by co-administration to the host of one or more immunosuppressants, such as cyclosporine, tacrolimus, sirolimus, cyclophosphamide, etc.

[0216] T lymphocytes, e.g., unmodified T lymphocytes, or T lymphocytes that express CD3 and CD28 or that contain a polypeptide comprising a CD3 zeta signaling domain and a CD28 costimulatory domain, can be expanded using antibodies against CD3 and CD28, e.g., antibodies attached to beads; see, e.g., U.S. Pat. Nos. 5,948,893; 6,534,055; 6,352,694; 6,692,964; 6,887,466; and 6,905,681.

[0217] Modified immune cells, such as modified T lymphocytes, may contain a "suicide gene" or "safety switch" that allows for the death of substantially all modified immune cells, if required. For example, modified T lymphocytes may contain the HSV thymidine kinase gene (HSV-TK), which in certain embodiments causes the death of modified T lymphocytes when contacted with ganciclovir. In another embodiment, modified T lymphocytes contain an inducible caspase, such as inducible caspase 9 (iCaspase 9), such as a fusion protein of caspase 9 with human FK506 binding protein that allows dimerization with certain small molecule drugs. See Straathof et al., Blood 105(11):4247-4254 (2005).

[0218] In certain embodiments, the inhibitor of NSD2 provided herein is administered to subjects with various types or stages of multiple myeloma in combination with chimeric antigen receptor (CAR) T cells.In certain embodiments, the CART cells in the combination target B cell maturation antigen (BCMA), and in more specific embodiments, the CAR T cells are bb2121 or bb21217.In some embodiments, the CAR T cells are JCARH125.

[0219] In certain embodiments, the inhibitor of NSD2 provided herein is administered to subjects with various types or stages of multiple myeloma in combination with a deacetylase inhibitor, such as a histone deacetylase inhibitor (HDAC inhibitor). Suitable DAC inhibitors or HDAC inhibitors include, for example, 1) hydroxamic acid derivatives; and 2) short chain fatty acids (SCFAs); 3) cyclic tetrapeptides; 4) benzamides; 5) electrophilic ketones; and / or any other type of compound that can inhibit histone deacetylase. In certain embodiments, the HDAC inhibitor is, for example, suberoylanilide hydroxamic acid (SAHA) or LAQ824. In certain embodiments, the HDAC is panobinostat. In certain embodiments, the HDAC inhibitor is panobinostat and is used in combination with bortezomib and dexamethasone.

[0220] In certain embodiments, the inhibitor of NSD2 provided herein is administered to subjects with various types or stages of multiple myeloma in combination with anti-SLAMF7 antibody.In certain embodiments, the anti-SLAMF7 antibody is elotuzumab.In certain embodiments, the anti-SLAMF7 antibody is elotuzumab and is used in combination with lenalidomide and dexamethasone.

[0221] In certain embodiments, the inhibitors of NSD2 provided herein are administered in combination with antibody drug conjugates, i.e., antibodies conjugated to a conjugation moiety or an agent such as a label or a toxin, to subjects with various types or stages of multiple myeloma. The conjugation moiety may be any conjugation moiety that would be useful to those skilled in the art. For example, the conjugation moiety may be a polymer, such as polyethylene glycol, that can improve the stability of the antibody in vitro or in vivo. The conjugation moiety may have a therapeutic activity, thereby resulting in an antibody drug conjugate. The conjugation moiety may be a molecular payload that is harmful to the target cell. The conjugation moiety may be a label that is useful for detection or diagnosis. In certain aspects, the conjugation moiety is linked to the antibody via a direct covalent bond. In certain aspects, the conjugation moiety is linked to the antibody via a linker. In certain aspects, the conjugation moiety or linker is attached via one or more unnatural amino acids of the antibody.

[0222] In certain embodiments, the inhibitor of NSD2 provided herein is administered to subjects with various types or stages of multiple myeloma in combination with a nuclear export inhibitor, also known as selective nuclear export inhibitor (SINE).In certain embodiments, the nuclear export inhibitor is leptomycin B.In certain embodiments, the nuclear export inhibitor is an exportin 1 (XPO1) inhibitor, such as selinexor (KPT-330).In certain embodiments, the nuclear export inhibitor is selinexor and is used in combination with dexamethasone.

[0223] It is understood that the above detailed description and the accompanying examples are illustrative only and should not be interpreted as limiting the scope of the present subject matter.Various changes and modifications to the embodiments of the present disclosure will be apparent to those skilled in the art.Such changes and modifications, including but not limited to those related to the chemical structures, substituents, derivatives, intermediates, synthesis, formulation and / or methods of use provided herein, can be made without departing from the spirit and scope thereof.The U.S. patents and publications referred to in this specification are incorporated by reference. EXAMPLES

[0224] Example 1. Analysis of a Newly Diagnosed Multiple Myeloma Patient Dataset Samples were obtained from 329 newly diagnosed multiple myeloma (NDMM) patients. Samples from the 329 NDMM patients were analyzed using either whole exome sequencing or whole genome sequencing. Each of the patient samples had a previously identified T4;14 translocation. Figure 1A shows the survival probability of the 329 NDMM patients over time.

[0225] A landmark analysis of 329 NDMM patients was then performed using overall survival of less than 24 months. Figure 1B shows a density plot of overall survival of 329 NDMM patients. A subset of high-risk patients (HR) was then identified and grouped by patients exhibiting an overall survival of less than 24 months (Figure 1C). MANTA output was parsed to extract translocation calls with a breakpoint at 4p, corresponding to the previously identified T4;14 translocation. Translocations between chromosomes 4 and 14 were extracted into data matrices containing the call, translocation partner position, variant allele frequency, and distance of the translocation from a fixed genomic point: the transcription start site of FGFR3. These data matrices were used for supervised analysis with a defined separation between high risk and non-high risk. Further genomic analysis was then performed on a subset of high-risk patient samples, which were samples derived from patients exhibiting an overall survival of less than 24 months.

[0226] Further genomic analysis demonstrated differences in overall survival that correlated with translocations that either disrupted the coding for the nuclear receptor-binding SET domain protein 2 (NSD2) protein or did not disrupt the coding for the NSD2 protein. As demonstrated in Figure 2A, translocations that disrupt the coding for the NSD2 protein correlated with shorter overall survival.

[0227] The patient data were then grouped into three regions with respect to the nuclear receptor binding SET domain protein 2 (NSD2) gene: (1) upstream of the transcription start site, (2) upstream of the canonical translation start site, and (3) within the gene body disrupting the coding sequence. Kaplan-Meier plots of the patient groups for these three regions were generated. Using these genomic features (canonical transcription and translation start sites), the patients were classified into three groups: no disruption, first half disruption, and second half disruption, with each group showing an increasingly poor prognosis in terms of overall survival (see Figure 2B). No disruption was defined as a T4:14 translocation with a breakpoint between nucleotides 1,808,872 and 1,871,393 on the forward strand according to genome build GRCh38.p13. First half disruption was defined as a T4:14 translocation with a breakpoint between nucleotides 1,871,393 and 1,900,655 on the forward strand according to genome build GRCh38.p13. The late disruption was defined as the T4:14 translocation with a breakpoint in the NSD2 gene after nucleotide 1,900,655 on the forward strand according to genome build GRCh38.p13. This example demonstrates that disruption of the NSD2 gene correlates with reduced overall survival in NDMM patients with the T4:14 translocation.

[0228] Example 2. Analysis of patient RNA-SEQ data Fusion transcripts arise from chromosomal rearrangements and are drivers of certain cancers, including multiple myeloma. NDMM patient RNA-seq data were analyzed with the STAR-fusion pipeline to identify expressed fusion transcripts. Fusions were found to occur either with the first NSD2 coding exon, generating full-length coding transcripts, or with later NSD2 coding exons, generating truncated transcripts. Analysis of the RNA-seq data identified three patient groups: (1) a full-length coding fusion transcript group with no disruption of the protein coding sequence or translated protein, (2) a truncated fusion transcript group characterized by late DNA breaks, and (3) a small subset of patients in whom fusion transcript expression was not measured. Kaplan-Meier plots were generated for these three patient groups based on fusion transcript expression. Patients expressing truncated fusion proteins had poorer overall survival than patients expressing full-length coding fusion transcripts or patients expressing no fusion transcripts (see Figure 3B).

[0229] The above written specification is believed to be sufficient for one skilled in the art to practice the embodiments. In the above description and examples, certain embodiments are detailed and the best mode contemplated by the inventors is explained. However, no matter how detailed the above contents are described in the text, it will be understood that the present embodiments can be practiced in many ways and should be construed according to the appended claims and any equivalents thereof.

Claims

1. A pharmaceutical composition for treating multiple myeloma, comprising a therapeutically effective amount of an inhibitor of nuclear receptor-binding SET domain protein 2 (NSD2).

2. 2. The method of claim 1, wherein the multiple myeloma has previously been determined to have a 4:14 chromosomal translocation (t(4;14)).

3. The pharmaceutical composition of claim 2, wherein the t(4;14) results in a disruption of the NSD2 gene.

4. The pharmaceutical composition of claim 3 , wherein the disruption of the NSD2 gene is located after the transcription start site of NSD2.

5. 5. The pharmaceutical composition of claim 3 or claim 4, wherein the disruption of the NSD2 gene is located after the translation start site of NSD2.

6. 6. The pharmaceutical composition of claim 3, wherein the disruption of the NSD2 gene is located before the translation stop site of NSD2.

7. 6. The pharmaceutical composition of claim 3, wherein the disruption of the NSD2 gene is located before the first coding exon of the NSD2 gene.

8. 7. The pharmaceutical composition of claim 3, wherein the disruption of the NSD2 gene is located in the first coding exon of the NSD2 gene.

9. 7. The pharmaceutical composition of claim 3, wherein the disruption of the NSD2 gene is located between the start of the first coding exon and the start of the second coding exon of the NSD2 gene.

10. 10. The pharmaceutical composition of any one of claims 3 to 9, wherein the disruption in the NSD2 gene is located at or after genomic position 1,871,393 of Genome Reference Consortium Human Build 38 Patch Release 13 (GRCh38.p13).

11. 11. The pharmaceutical composition of any one of claims 3 to 10, wherein the disruption of the NSD2 gene is located between genomic position 1,871,393 and genomic position 1,900,655 of GRCh38.p13.

12. 11. The pharmaceutical composition of any one of claims 3 to 10, wherein the disruption of the NSD2 gene is located at or after genomic position 1,900,655 of GRCh38.p13.

13. 11. The pharmaceutical composition of any one of claims 3 to 10, wherein the t(4;14) is located between genomic position 1,900,655 and genomic position 1,982,207 of GRCh38.p13.

14. 14. The pharmaceutical composition of any one of claims 1 to 13, wherein the multiple myeloma expresses a truncated NSD2 protein.

15. 14. The pharmaceutical composition of any one of claims 1 to 13, wherein the multiple myeloma expresses a full-length NSD2 protein.

16. 16. The pharmaceutical composition of claim 15, wherein the multiple myeloma expresses high levels of full-length NSD2 protein.

17. 17. The pharmaceutical composition of any one of claims 2 to 16, wherein the 4:14 chromosomal translocation (t(4;14)) has been identified by a method comprising in situ hybridization, PCR, RT-PCR, RNA sequencing, fluorescent in situ hybridization (FISH), transcript in situ hybridization, whole genome sequencing, whole exome sequencing, mixed ligation probe assay, mass spectrometry, and / or MALDI-TOF.

18. 18. The pharmaceutical composition of any one of claims 1 to 17, wherein the inhibitor of NSD2 is selected from an antibody, a small molecule, an aptamer, an siRNA, and an antisense oligonucleotide.

19. 19. A pharmaceutical composition according to any one of claims 1 to 18, characterized in that it is used to administer at least one second therapeutic agent.

20. 20. The pharmaceutical composition of claim 19, wherein the at least one second therapeutic agent is selected from a chemotherapeutic agent, a steroid, an immunomodulatory agent, a proteasome inhibitor, a histone deacetylase inhibitor, an anti-CD38 antibody, an anti-SLAMF7 antibody, an antibody drug conjugate, and a nuclear export inhibitor.

21. 20. The pharmaceutical composition of claim 19, wherein the at least one second therapeutic agent is selected from lenalidomide, thalidomide, pomalidomide, bortezomib, carfilzomib, ixazomib, panobinostat, melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, bendamustine, dexamethasone, prednisone, daratumumab, isatuximab, elotuzumab, belantamab mafodotin-blmf, selinexor, pamidronate, zoledronic acid, and denosumab.

22. The at least one second therapeutic agent is a) lenalidomide; b) iberdomide; c) (S)-4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)methyl)benzyl)piperazin-1-yl)-3-fluorobenzonitrile; d) (i) lenalidomide, pomalidomide, or thalidomide; and (ii) dexamethasone; e) (i) carfilzomib, ixazomib, or bortezomib; (ii) lenalidomide; and (iii) dexamethasone; f) (i) bortezomib or carfilzomib; (ii) cyclophosphamide; and (iii) dexamethasone; g) (i) elotuzumab or daratumumab; (ii) lenalidomide; and (iii) dexamethasone; h) bortezomib, liposomal doxorubicin, and dexamethasone; i) panobinostat, bortezomib, and dexamethasone; j) elotuzumab, bortezomib, and dexamethasone; k) melphalan and prednisone with or without thalidomide or bortezomib; l) Vincristine, doxorubicin, and dexamethasone; m) dexamethasone, cyclophosphamide, etoposide, and cisplatin; and n) Dexamethasone, thalidomide, cisplatin, doxorubicin, cyclophosphamide, and etoposide, with or without bortezomib 20. The pharmaceutical composition of claim 19, wherein the pharmaceutical composition is selected from:

23. A pharmaceutical composition for treating multiple myeloma comprising an NSD2 inhibitor, the pharmaceutical composition being used to determine whether a subject having multiple myeloma has a 4:14 chromosomal translocation (t(4;14)), and if the subject has the t(4;14) translocation, to select the subject for treatment with the NSD2 inhibitor, and to administer the NSD2 inhibitor to the subject.

24. A pharmaceutical composition for treating multiple myeloma comprising an NSD2 inhibitor, characterized in that the pharmaceutical composition is used to determine whether a subject having multiple myeloma has a 4:14 chromosomal translocation (t(4;14)), and if the subject has the t(4;14) translocation, the subject is predicted to benefit from treatment with an NSD2 inhibitor, and the NSD2 inhibitor is administered to the subject.

25. 25. The pharmaceutical composition of claim 23 or claim 24, wherein the t(4;14) results in a disruption of the NSD2 gene.

26. 26. The pharmaceutical composition of claim 25, wherein the disruption of the NSD2 gene is located after the transcription start site of NSD2.

27. 27. The pharmaceutical composition of claim 25 or claim 26, wherein the disruption of the NSD2 gene is located after the translation start site of NSD2.

28. 28. The pharmaceutical composition of any one of claims 25 to 27, wherein the disruption of the NSD2 gene is located before the translation stop site of NSD2.

29. 29. The pharmaceutical composition of any one of claims 25 to 28, wherein the disruption of the NSD2 gene is located before the first coding exon of the NSD2 gene.

30. 29. The pharmaceutical composition of any one of claims 25 to 28, wherein the disruption of the NSD2 gene is located in the first coding exon of the NSD2 gene.

31. 29. The pharmaceutical composition of any one of claims 25 to 28, wherein the disruption of the NSD2 gene is located between the start of the first coding exon and the start of the second coding exon of the NSD2 gene.

32. 32. The pharmaceutical composition of any one of claims 25 to 31, wherein the disruption in the NSD2 gene is located at or after genomic position 1,871,393 of Genome Reference Consortium Human Build 38 Patch Release 13 (GRCh38.p13).

33. 33. The pharmaceutical composition of any one of claims 25 to 32, wherein the disruption of the NSD2 gene is located between genomic position 1,871,393 and genomic position 1,900,655 of GRCh38.p13.

34. 33. The pharmaceutical composition of any one of claims 25 to 32, wherein the disruption of the NSD2 gene is located at or after genomic position 1,900,655 of GRCh38.p13.

35. 33. The pharmaceutical composition of any one of claims 25 to 32, wherein the t(4;14) is located between genomic position 1,900,655 and genomic position 1,982,207 of GRCh38.p13.

36. 36. The pharmaceutical composition of any one of claims 23 to 35, wherein the multiple myeloma expresses a truncated NSD2 protein.

37. 36. The pharmaceutical composition of any one of claims 23 to 35, wherein the multiple myeloma expresses a full-length NSD2 protein.

38. 38. The pharmaceutical composition of claim 37, wherein the multiple myeloma expresses high levels of full-length NSD2 protein.

39. 39. The pharmaceutical composition of any one of claims 23 to 38, wherein determining whether the subject has t(4;14) comprises in situ hybridization, PCR, RT-PCR, RNA sequencing, fluorescent in situ hybridization (FISH), transcript in situ hybridization, whole genome sequencing, whole exome sequencing, mixed ligation probe assay, mass spectrometry, and / or MALDI-TOF.

40. 40. The pharmaceutical composition of any one of claims 23 to 39, wherein the inhibitor of NSD2 is selected from an antibody, a small molecule, an aptamer, an siRNA, and an antisense oligonucleotide.

41. 41. The pharmaceutical composition of any one of claims 23 to 40, further comprising administering at least one second therapeutic agent.

42. 42. The pharmaceutical composition of claim 41, wherein the at least one second therapeutic agent is selected from a chemotherapeutic agent, a steroid, an immunomodulatory agent, a proteasome inhibitor, a histone deacetylase inhibitor, an anti-CD38 antibody, an anti-SLAMF7 antibody, an antibody drug conjugate, and a nuclear export inhibitor.

43. 42. The pharmaceutical composition of claim 41, wherein the at least one second therapeutic agent is selected from lenalidomide, thalidomide, pomalidomide, bortezomib, carfilzomib, ixazomib, panobinostat, melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, bendamustine, dexamethasone, prednisone, daratumumab, isatuximab, elotuzumab, belantamab mafodotin-blmf, selinexor, pamidronate, zoledronic acid, and denosumab.

44. The at least one second therapeutic agent is a) lenalidomide; b) iberdomide; c) (S)-4-(4-(4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)methyl)benzyl)piperazin-1-yl)-3-fluorobenzonitrile; d) (i) lenalidomide, pomalidomide, or thalidomide; and (ii) dexamethasone; e) (i) carfilzomib, ixazomib, or bortezomib; (ii) lenalidomide; and (iii) dexamethasone; f) (i) bortezomib or carfilzomib; (ii) cyclophosphamide; and (iii) dexamethasone; g) (i) elotuzumab or daratumumab; (ii) lenalidomide; and (iii) dexamethasone; h) bortezomib, liposomal doxorubicin, and dexamethasone; i) panobinostat, bortezomib, and dexamethasone; j) elotuzumab, bortezomib, and dexamethasone; k) melphalan and prednisone, with or without thalidomide or bortezomib; l) Vincristine, doxorubicin, and dexamethasone; m) dexamethasone, cyclophosphamide, etoposide, and cisplatin; and n) Dexamethasone, thalidomide, cisplatin, doxorubicin, cyclophosphamide, and etoposide, with or without bortezomib 42. The pharmaceutical composition of claim 41, wherein said pharmaceutical composition is selected from:

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