Chimeric antigen receptor t cell therapy

Abstract

To provide methods of CAR-T cancer treatment and methods of predicting clinical outcomes in response to those treatments.SOLUTION: The present invention provides a method of predicting a response to CD19 CAR-T treatment in a subject having cancer, comprising measuring a baseline SPD in the subject, and determining the SPD range, wherein a low SPD range indicates a likelihood of positive response to the CAR-T treatment.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present application relates to methods of CAR-T cancer treatments and methods of predicting clinical outcomes for those treatments. [Background technology]

[0002] Immunotherapy cancer treatment relies on enriched or engineered human T cells to target and kill cancer cells in patients. To increase the ability of T cells to target and kill specific cancer cells, methods have been developed to engineer T cells to express constructs that direct T cells to specific target cancer cells. Chimeric antigen receptors (CARs) and engineered T cell receptors (TCRs), which contain binding domains that can interact with specific tumor antigens, enable T cells to target and kill cancer cells that express specific tumor antigens. Adoptive cell therapy targeting CD19-expressing cancer cells has shown promising results with excellent clinical outcomes. There is a need to predict clinical outcomes for CAR-T therapy. Summary of the Invention

[0003] The present application relates to methods of CAR-T cancer treatments and methods of predicting clinical outcomes to those treatments. Methods of predicting potential clinical outcomes include using the baseline sum of product diameters (SPD) of index lesions and / or the number of prior lines of therapy at baseline as indicators.

[0004] It is to be understood that the disclosure is not limited in its application to the details set forth in the following embodiments, claims, detailed description and drawings, as the disclosure is capable of other embodiments and of being practiced or carried out in many other ways.

[0005] Below are some exemplary embodiments of the present disclosure.

[0006] Embodiment 1. A method of predicting a response to CD19 CAR-T therapy in a subject having cancer, comprising measuring baseline SPD in the subject and determining an SPD range, wherein a low SPD range indicates a likelihood of a positive response to the CAR-T therapy.

[0007] Embodiment 2. The method of embodiment 1, wherein the SPD can fall into one of four ranges, where the lower the range the SPD falls into, the higher the likelihood of response to CD19 CAR-T therapy.

[0008] Embodiment 3. The four ranges are as follows: SPD quartile 1, approximately 100 mm 2 (Including) ~ approx. 2000mm 2 (including), approx. 840mm 2 Median SPD of SPD quartile 2, approximately 2000 mm 2 (excluding) ~ approx. 3700mm 2 (including), approx. 2820mm 2 Median SPD of SPD quartile 3, approximately 3700 mm 2 (excluding) ~ approx. 6700mm 2 (including), approx. 5100mm 2 The median SPD of, and SPD quartile 4, approximately 6700 mm 2 (not included) ~ approx. 24000mm 2 (including), approx. 9300mm 2 Median SPD of The method of embodiment 2, comprising:

[0009] Embodiment 4. If the SPD value at baseline is in the SPD quartile 1 to 4, In the event that the subject is subsequently treated with a CD19 CAR-T therapy, the method of any one of embodiments 1-3.

[0010] Embodiment 5. A method of treating cancer in a subject in need of treatment, comprising administering a therapeutically effective amount of CD19 to a subject having a baseline SPD value in the SPD quartiles 1-4. administering a CAR-T therapeutic agent.

[0011] Embodiment 6. A method for predicting the likelihood of relapse after CD19 CAR-T therapy in a subject having cancer, comprising determining the number of prior lines of therapy in the subject and determining into which of four ranges the number falls, wherein a higher number of prior lines of therapy predicts a higher likelihood of relapse after CD19 CAR-T therapy for the subject.

[0012] Embodiment 7. A method for predicting the likelihood of a sustained response to CD19 CAR-T therapy in a subject with cancer, comprising measuring the number of prior lines of therapy in the subject at baseline and determining in which of four ranges the number falls, wherein a lower range indicates a likelihood of a sustained response to CD19 CAR-T therapy.

[0013] Embodiment 8. The method of embodiment 6 or 7, wherein the number of pretreatment lines is in the range of 1 to 2, 3, 4 or 5 or more.

[0014] Embodiment 9. The method of any one of embodiments 6 to 8, further comprising subsequently administering a CD19 CAR-T therapy to the subject who has received 1 to 2, 3, 4, or 5 or more lines of prior therapy.

[0015] Embodiment 10. A method of treating cancer in a subject in need of treatment, comprising administering a therapeutically effective amount of a CD19 CAR-T therapy to a subject, the subject having had 1 to 2, 3, 4, or 5 or more lines of prior therapy.

[0016] Embodiment 11. The method of any one of embodiments 1 to 10, wherein the cancer is a hematological cancer or relapsed / refractory diffuse large B-cell lymphoma.

[0017] Embodiment 12. The method of any one of embodiments 1 to 11, wherein the CD19 CAR-T treatment comprises treatment with axicabtagene ciloleucel (Yescarta), tisagenlecleucel (Kymriah), JCAR017, JCAR015, JCAR014, Uppsala University anti-CD19 CAR (NCT02132624) or UCART19.

[0018] Embodiment 13. A method for predicting long-term durability of response to treatment of cancer with anti-CD19 CAR-T cell therapy in a patient in need of treatment, comprising assessing progression-free survival three months after a single administration of the therapeutic agent, wherein achievement of a complete or partial response at three months is predictive of long-term durability of response in said patient.

[0019] Embodiment 14. The method of embodiment 13, wherein the anti-CD19 CAR-T treatment comprises treatment with axicabtagene ciloleucel (Yescarta), tisagenlecleucel (Kymriah), JCAR017, JCAR015, JCAR014, Uppsala University anti-CD19 CAR (NCT02132624) or UCART19.

[0020] Embodiment 15. The method of embodiment 13 or 14, wherein the cancer is a blood cancer.

[0021] Embodiment 16. The method of embodiment 15, wherein the cancer is relapsed / refractory diffuse large B-cell lymphoma.

[0022] Embodiment 17. The method of any one of embodiments 13-16, wherein the long-term response durability includes a complete or partial response lasting for more than 9 months, more than 12 months, more than 18 months, or more than 24 months.

[0023] Embodiment 18. The method of any one of embodiments 1 to 17, wherein the subject is 65 years of age or older.

[0024] Embodiment 19. The method of any one of embodiments 1 to 17, wherein the subject is under 65 years of age.

[0025] Embodiment 20. The method of any one of embodiments 1 to 19, wherein the CD19 CAR-T therapeutic is administered as a first-line therapy. [Brief description of the drawings]

[0026] [Figure 1] FIG. 1 shows patient outcomes according to quartiles of SPD: AUC0-28 (area under the curve from day 0 to day 28), CRS (cytokine release syndrome), ORR (objective response rate), Q (quartiles). [Diagram 2] Figure 1 shows a post-hoc analysis of site-assessed progression-free survival by response status at 3 months after axicabtagene-ciloleucel. Sixty patients with ongoing complete response, partial response, or stable disease at 3 months of phase 2 are shown. The x-axis shows time from chimeric antigen receptor T-cell infusion. Four of eight patients with partial response and four of nine patients with stable disease at 3 months later converted to complete response. NR=not reached. NE=not estimable. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] Unless otherwise specified, scientific and technical terms used in connection with this disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art.

[0028] As used in this specification and the appended claims, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise. Unless specifically stated or apparent from the context, the term "or" as used herein is understood to include and encompass both "or" and "and." As used herein, the term "and / or" is understood as a specific disclosure of each of the two specified features or components with or without the other.

[0029] As described herein, any concentration range, percentage range, ratio range, or integer range is understood to include any integer value within the stated range, and, where appropriate, fractions thereof (such as tenths and hundredths of an integer), unless otherwise indicated. Unless specifically stated or clear from the context, the term "about" as used herein refers to a value or composition that falls within an acceptable error range for a particular value or composition as determined by one of ordinary skill in the art, depending in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "essentially composed of" can mean within one or more standard deviations per one practice in the art. "About" or "essentially composed of" can mean within a range of up to 10% (i.e., ±10%). Thus, "about" refers to a range that is within 10% of the indicated value. , 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, or 0.001% greater or less than the range. For example, about 5 mg can include any amount between 4.5 mg and 5.5 mg. Additionally, particularly with respect to biological systems or processes, the above terms can be understood to include up to 10 times a value (up It may mean up to an order of magnitude or up to five times the specific value or composition. As provided in this disclosure, unless otherwise indicated, the meaning of "about" or "consisting essentially of" is intended to include an acceptable range of error for that particular value or composition.

[0030] The term "administering" is known to those skilled in the art. Refers to the physical introduction of an agent into a subject using any of a variety of methods and delivery systems. Exemplary routes of administration for the formulations disclosed herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes of administration, such as by injection or infusion. As used herein, the phrase "parenteral administration" refers to a mode of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intraarticular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subarticular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. In some embodiments, the formulations are administered by a route other than parenteral, such as orally. Other non-parenteral routes include topical, epidermal, or mucosal routes of administration, such as intranasal, vaginal, rectal, sublingual, or topical. Administration can also be, for example, once, multiple times, and / or over one or more extended periods of time. In some embodiments, administration is by infusion. For example, an infusion bag of a CD19-directed genetically modified autologous T-cell immunotherapeutic contains approximately 68 mL of a suspension of chimeric antigen receptor (CAR)-positive T cells. The target dose is approximately 1×10 per kg of body weight. 6 Approximately 2 x 10 pieces 6 Between 10 and 2 x 10 CAR-positive viable T cells 8 In some embodiments, the CD19-directed genetically modified autologous T cell immunotherapeutic is Axi-cel™ (YESCARTA™, axi-cbutagen-ciloleucel).

[0031] The term "lymphocyte" as used herein includes natural killer (NK) cells, T cells, or B cells. NK cells are a type of cytotoxic (cell-toxic) lymphocyte that constitutes the main component of the innate immune system. NK cells reject tumor and virus-infected cells. NK cells act by the process of apoptosis or programmed cell death. NK cells are named "natural killer" because they do not require activation to kill cells. T cells play a major role in cell-mediated immunity (not involving antibodies). Their T cell receptor (TCR) differentiates them from other types of lymphocytes. The thymus, a specialized organ of the immune system, is primarily responsible for the maturation of T cells. There are six types of T cells: helper T cells (e.g., CD4 + cell), cytotoxic T cell (TC, cytotoxic T lymphocyte, CTL, T killer cell, cytolytic T cell, CD8 + (i) stem cell memory T cells, also known as T cells or killer T cells, such as naive T cells, express CD45RO - , CCR7 + , CD45RA + , CD62L + (L-selectin), CD27 + , CD28 + and IL-7Rα + However, they also express large amounts of CD95, IL-2Rβ, CXCR3 and LFA-1, and exhibit many functional attributes characteristic of memory cells; (ii) central memory T CM (iii) effector memory TEM cells express L-selectin and CCR7 and secrete IL-2 but not IFNγ or IL-4; and (iv) effector memory TEM cells do not express L-selectin or CCR7 but produce effector cytokines such as IFNγ and IL-4), regulatory T cells (Tregs, suppressor T cells or CD4 + CD25 +There are three types of T cells: regulatory T cells (Treg), natural killer T cells (NKT) and gamma delta T cells. On the other hand, B cells play the most important role in humoral immunity (antibody-mediated immunity). B cells produce antibodies and antigens, act as antigen-presenting cells (APCs), and, after activation by antigen interaction, mediate the immune response to antigens. In mammals, immature B cells are formed in the bone marrow, from which they get their name.

[0032] As used herein, a "patient" includes any human afflicted with a disease or condition, such as cancer (e.g., lymphoma or leukemia). The terms "subject" and "patient" are used interchangeably herein.

[0033] The term "immunotherapy" refers to the treatment of a subject afflicted with a disease or at risk of contracting a disease or suffering from a recurrence of a disease by methods that involve inducing, enhancing, suppressing, or ameliorating an immune response. Examples of immunotherapy include, but are not limited to, T cell therapy. T cell therapy includes adoptive T cell therapy, tumor infiltrating lymphoma therapy, and immunotherapy. The conditioning methods may include T-cell immunotherapy, autologous cell therapy, engineered autologous cell therapy (eACT™), and allogeneic T-cell transplantation. One skilled in the art will recognize that the conditioning methods disclosed herein may enhance the efficacy of any transplanted T-cell therapy. Examples of T-cell therapy are described in U.S. Patent Application Publication Nos. 2014 / 0154228 and 2002 / 0006409, U.S. Patent No. 5,728,388, and WO 2008 / 081035. Other non-limiting examples can be found in U.S. Patent No. 9,855,298, U.S. Patent Application Publication Nos. 20190151361, 20190144515, 20190093101, 20190032011, 20190092818, 20180369283, 20180296601, 20180280437, 20180230224, 20180086846, and 20180016620. Exemplary reviews of T cell therapies used in the art include Jafferji MS, Yang JC, Adoptive T-Cell Therapy for Solid Malignancies, Surg Oncol Clin N Am. 2019 Jul;28(3):465-479. doi: 10.1016 / j.soc.2019.02.012. Epub 2019 Apr 12, Minutolo NG, Hollander EE, Powell DJ Jr.. The Emergence of Universal Immune Receptor T Cell Therapy for Cancer, Front Oncol. 2019 Mar 26;9:176. doi: 10.3389 / fonc.2019.00176. eCollection 2019, Strati P, Neelapu SS., Chimeric Antigen Receptor-Engineered T Cell Therapy in Lymphoma, Curr Oncol Rep. 2019 Mar 27;21(5):38. doi: 10.1007 / s11912-019-0789-z.

[0034] The term "autologous" refers to any material derived from the same individual that is subsequently reintroduced into the same individual. For example, the autologous cell therapy (ACT) method described herein involves collecting lymphocytes from a patient, then engineering the lymphocytes to express, for example, a CAR construct, and then administering them back to the same patient. The term "autologous cell therapy," which can be abbreviated as "ACT" and is also known as adoptive cell transfer, is the process of collecting a patient's own T cells and then genetically modifying them to recognize and target one or more antigens expressed on the cell surface of one or more specific tumor cells or malignancies.

[0035] "Therapeutically effective" means that the use of CD19 CAR-T to treat a patient's cancer results in some demonstrated clinical benefit compared to no treatment (if appropriate) or known standard treatment. Clinical benefit in a patient population can be assessed by any method known to those skilled in the art. In one embodiment, clinical benefit can be assessed based on objective response rate (ORR), duration of response (DOR), progression-free survival (PFS), sustained response at 1 year, and / or overall survival (OS). Objective response rate (ORR) is defined as the proportion of participants who achieve a complete response (CR) or partial response (PR).

[0036] In some embodiments, a complete response indicates therapeutic benefit. In some embodiments, a partial response indicates therapeutic benefit. In some embodiments, stable disease indicates therapeutic benefit. In some embodiments, an increase in overall survival indicates therapeutic benefit. In some embodiments, therapeutic benefit may constitute an improvement in time to disease progression and / or an improvement in symptoms or quality of life. In other embodiments, the therapeutic benefit does not translate into increased duration of disease control, but instead, a significant reduction in symptom burden resulting in improved quality of life.

[0037] Chimeric antigen receptors (CARs) The chimeric antigen receptors (CAR or CAR-T) and T cell receptors (TCR) of the present disclosure are genetically engineered receptors. These engineered receptors can be easily inserted into immune cells, including T cells, by techniques known in the art and expressed by the cells. In CARs, a single receptor can be programmed to recognize a specific antigen and, when bound to the antigen, activate the immune cell to attack and destroy cells that carry or express the antigen. If these antigens are present on tumor cells, the immune cells expressing the CAR can target and kill the tumor cells.

[0038] One aspect of the present invention is a chimeric antigen receptor (CAR) or T cell receptor comprising (i) an antigen binding molecule, (ii) a costimulatory domain, and (iii) an activation domain. The costimulatory domain may comprise an extracellular domain, a transmembrane domain, and an intracellular domain. In some embodiments, the extracellular domain comprises a hinge domain or a truncated hinge domain.

[0039] In some embodiments, an antigen-binding molecule is a molecule that comprises the antigen-binding portion (e.g., CDR) of an antibody from which it is derived. An antigen-binding molecule may comprise an antigen complementarity determining region (CDR). Examples of antigen-binding molecules include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, dAbs, linear antibodies, scFv antibodies, and multispecific antibodies formed from antigen-binding molecules. Peptibodies (i.e., Fc fusion molecules containing a peptide binding domain) are another example of a suitable antigen-binding molecule. In one embodiment, a CD19 CAR construct comprises an anti-CD19 single chain Fv. A "single chain Fv" or "scFv" antibody-binding fragment comprises the variable heavy (V) domain of an antibody. H ) domain and variable light (V L ) domains, which are present in a single polypeptide chain. Generally, an Fv polypeptide comprises a V H Domain and V LThe scFv further comprises a polypeptide linker between the domains which enables the scFv to form the desired structure for antigen binding. All antibody-related terms used herein take on their conventional meaning in the art and are well understood by those of skill in the art.

[0040] In some embodiments, the CAR comprises one or more costimulatory domains. In some embodiments, the costimulatory domains are selected from the group consisting of CD28, OX-40, 4-1BB / CD137, CD2, CD7, CD27, CD30, CD40, programmed death-1 (PD-1), inducible T cell costimulatory molecule (ICOS), lymphocyte function associated antigen-1 (LFA-1 (CD11a / CD18)), CD3 gamma, CD3 delta, CD3 epsilon, CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), NKG2C, Ig alpha (CD79a), DAP-10, Fc gamma receptor, MHC class I molecule, TNF receptor protein, immunoglobulin-like protein, cytokine receptor, integrin, signaling lymphocyte activation molecule (SLAM protein), activating NK cell receptor, BTLA, Toll ligand receptor, ICAM-1, B7- H3, CDS, ICAM-1, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8 alpha, CD8 beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6 , CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, C D18, LFA-1, ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1(CD226), SLAMF4(CD244, 2B4), CD84, CD96(Tactile), The signal transduction domain of a ligand that specifically binds to CEACAM1, CRT AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83, or any combination thereof.

[0041] In some embodiments, the intracellular domain is selected from the group consisting of 4-1BB / CD137, activating NK cell receptor, B7-H3, BAFFR, BLAME (SLAMF8), BTLA, CD100 (SEMA4D), CD103, CD160 (BY55), CD18, CD19, CD19a, CD2, CD247, CD27, CD276 (B7-H3), CD29, CD3 delta, CD3 epsilon, CD3 gamma, CD30, CD4, CD40, CD49a, CD49D, CD49f, CD69, CD7, CD84, CD8 alpha, CD8 beta, CD96 (Tactile), CD11a, CD11b, CD11c, CD11d, CDS, CEACAM1, CRT AM, cytokine receptor, DAP-10, DNAM1 (CD226), Fc gamma receptor, GADS, GITR, HVEM (LIGHTR), IA4, ICAM-1, ICAM-1, Ig alpha (CD79a), IL2R beta, IL2R gamma, IL7R alpha, immunoglobulin-like protein, inducible T cell costimulator (ICOS), integrin, ITGA4, ITGA4, ITGA6, ITGAD, ITGAE, ITGAL, ITGAM, ITGAX, ITGB2, ITGB7, ITGB1, KIRDS2, LAT, LFA-1, LFA-1, ligand that specifically binds to CD83, LIGHT, LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), LTBR, ​​Ly9 (CD2 29), lymphocyte function associated antigen-1 (LFA-1 (CD11a / CD18)), MHC class I molecule, NKG2C, NKG2D, NKp30, NKp44, NKp46, NKp80 (KLRF1), OX-40, PAG / Cbp, programmed death-1 (PD-1), PSGL1, SELPLG (CD162), signaling lymphocyte activation molecule (SLAM protein), SLAM (SLAMF1; CD150; IPO-3), SLAMF4 (CD244; 2B4), SLAMF6 (NTB-A; Ly108), SLAMF7, SLP-76, TNF receptor protein, TNFR2, Toll ligand receptor, TRANCE / RANKL, signaling region of VLA1 or VLA-6, or combinations thereof. In some embodiments, the CAR comprises a hinge region between the transmembrane domain and the binding molecule.In some embodiments, the hinge region is an IgG1, IgG2, IgG3, IgG4, IgA, IgD, IgE, IgM, CD28, or CD8 alpha hinge region.

[0042] In some embodiments, the transmembrane domain is a transmembrane domain of CD28, 4-1BB / CD137, the alpha chain of the T cell receptor, the beta chain of the T cell receptor, CD3 epsilon, CD4, CD5, CD8 alpha, CD9, CD16, CD19, CD22, CD33, CD37, CD45, CD64, CD80, CD86, CD134, CD137, CD154, or the zeta chain of the T cell receptor, or any combination thereof. In some embodiments, the activation domain can be derived, for example, from any form of CD3 zeta. In some embodiments, the activation domain is derived from DAP10, DAP12, or other TCR-type activation signaling molecules.

[0043] In one aspect, the present application relates to CD19 CAR T cell therapy. In one embodiment, the CD19 CAR construct comprises an anti-CD19 scFv domain, an intracellular domain, a transmembrane domain, a costimulatory domain, and an activation domain. In one embodiment, the transmembrane domain is derived from the transmembrane domain of CD28, 4-1BB / CD137, CD8 alpha, or any combination thereof. In one embodiment, the costimulatory domain is derived from CD8, CD28 OX40, 4-1BB / CD137, or a combination thereof. In an embodiment, the activation domain is derived from CD3 zeta. In one embodiment, the CD19 CAR construct comprises a 4-1BB costimulatory domain. In one embodiment, the CD19 The CAR construct comprises a CD28 costimulatory domain. In one embodiment, the CAR construct comprises a CD19 The CAR construct comprises an anti-CD19 scFv, a hinge / transmembrane domain and a costimulatory domain from CD28, and an activation domain from CD3 zeta. In one embodiment, the CAR is an axicabtagene ciloleucel-expressed CAR. In one embodiment, the CAR is a Kymriah™-expressed CAR. Additional CD19-directed CARs that can be used in the methods of the disclosure include, but are not limited to, JCAR017, JCAR015, JCAR014, Uppsala University anti-CD19 CAR (NCT02132624), and UCART19 (Cellectis). Sadelain et al. See al. Nature Rev. Cancer Vol. 3 (2003), Ruella et al., Curr Hematol Malig Rep., Springer, NY (2016), and Sadelain et al. Cancer Discovery (Apr 2013). .

[0044] CAR-T cells The T cells of the immunotherapy may be engineered to express any of the above CARs or others and are referred to as CAR-T cells. The CAR-T cells may also be engineered to express other molecules and may be any one of the following exemplary types or others available in the art: first generation, second generation, third generation, fourth generation, fifth generation (etc.) CAR-T cells, armed CAR-T cells, motile CAR-T cells, TRUCK T cells, switch receptor CAR-T cells, gene edited CAR T cells, dual receptor CAR T cells, suicide CAR T cells, drug inducible CAR-T cells, synNotch inducible CAR T cells, and suppressive CAR T cells.

[0045] T cells for immunotherapy can be derived from any source known in the art. For example, T cells can be differentiated in vitro from a hematopoietic stem cell population, or T cells can be obtained from a subject. T cells can be obtained, for example, from peripheral blood mononuclear cells (PBMCs), bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from an infection site, ascites, pleural effusion, splenic tissue, and tumors. Additionally, T cells can be derived from one or more T cell lineages available in the art. T cells can also be obtained from a unit of blood collected from a subject using any number of techniques known to those skilled in the art, such as FICOLL™ separation and / or apheresis. Further methods of isolating T cells for T cell therapy are disclosed in US Patent Application Publication No. 2013 / 0287748. Other non-limiting examples can be found in International Application No. PCT / US2015 / 014520 (published as WO 2015 / 120096) and International Application No. PCT / US2016 / 057983 (published as WO 2017 / 070395), all of which are incorporated herein by reference in their entirety for purposes of describing these methods.

[0046] In one embodiment, the T cells are autologous T cells. In one embodiment, the T cells are autologous stem cells (for autologous stem cell therapy or ASCT). In one embodiment, the T cells are non-autologous T cells. In some embodiments, the T cells are obtained from a donor subject. In some embodiments, the donor subject is a human patient suffering from cancer or a tumor. In some embodiments, the donor subject is a human patient not suffering from cancer or a tumor. In some embodiments, the donor T cells used in T cell therapy are obtained from a patient (e.g., for autologous T cell therapy). In other embodiments, the donor T cells used in T cell therapy are obtained from a subject that is not a patient.

[0047] CD19 CAR-T cells can be used in a variety of applications, including, but not limited to, those described in International Application No. PCT / US2015 / 014520 (published as WO 2015 / 120096) and International Application No. PCT / US2016 / 057983 (published as WO 2017 / 070395). The T cells may be made by any method for producing T cells for immunotherapy, including the methods described in US Pat. No. 6,393,366 (both of which are incorporated herein by reference in their entireties for the purposes of describing these methods), any and all methods used to make axicabtagene ciloleucel or Yescarta™, any and all methods used to make tisagenlecleucel / Kymriah™, any and all methods used to make "off the shelf" T cells for immunotherapy, and any other method for making lymphocytes for administration to humans.

[0048] In one embodiment, the T cells may be obtained from, for example, peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. Additionally, the T cells may be derived from one or more T cell lineages available in the art. The T cells may also be obtained from a unit of blood collected from a subject using any number of techniques known to those skilled in the art, such as FICOLL™ separation and / or apheresis. In some embodiments, the cells collected by apheresis are washed to remove the plasma fraction and placed into a buffer or medium appropriate for further processing. In some embodiments, the cells are washed with PBS. As will be appreciated, a washing step may be used, for example, by using a semi-automated flow-through centrifuge such as the Cobe™ 2991 cell processing device, Baxter CytoMate™, etc. In some embodiments, the washed cells are resuspended in one or more biocompatible buffers or other saline solutions with or without buffers. In some embodiments, undesirable components of the apheresis sample are removed. Additional methods of isolating T cells for T cell therapy are disclosed in U.S. Patent Application Publication No. 2013 / 0287748, which is incorporated by reference in its entirety.

[0049] In some embodiments, T cells are isolated from PBMCs by lysis of red blood cells and depletion of monocytes, for example, by centrifugation through a PERCOLL™ gradient.+ , CD8 + , CD28 + , CD45RA + and CD45RO + Specific subpopulations of T cells, such as T cells from the negative selection group, can be further isolated by positive or negative selection techniques known in the art. For example, enrichment of a T cell population by negative selection can be accomplished by a combination of antibodies against surface markers unique to the cells to be negatively selected. In some embodiments, negative magnetic immunoadherence or flow cytometric sorting and / or selection of cells using a cocktail of monoclonal antibodies against cell surface markers present on the cells to be negatively selected can be used. For example, negative selection can result in the deletion of CD4 + To enrich for cells, the monoclonal antibody cocktail typically includes antibodies against CD8, CD11b, CD14, CD16, CD20, and HLA-DR. In some embodiments, flow cytometry and cell sorting are used to isolate cell populations of interest for use in the present disclosure.

[0050] In some embodiments, PBMCs are used directly for genetic modification with immune cells (such as CARs) using the methods described herein. In some embodiments, after isolating PBMCs, T lymphocytes are further isolated and sorted into naive, memory, and effector T cell subpopulations, either before or after genetic modification and / or expansion, both cytotoxic and helper T lymphocytes.

[0051] In some embodiments, CD8 + The cells are +By identifying cell surface antigens associated with the cells, they are further sorted into naive cells, central memory cells, and effector cells. In some embodiments, expression of phenotypic markers of central memory T cells includes expression of CCR7, CD3, CD28, CD45RO, CD62L, and CD127, and are negative for granzyme B. In some embodiments, central memory T cells are negative for CD8 + , CD45RO + and CD62L + T cells In some embodiments, effector T cells are negative for CCR7, CD28, CD62L, and CD127, and positive for granzyme B and perforin. + T cells can be further sorted into subpopulations. For example, CD4 + Helper T cells can be sorted into naive cells, central memory cells, and effector cells by identifying cell populations that have cell surface antigens.

[0052] T cells can be engineered to express, for example, a chimeric antigen receptor (CAR) or a T cell receptor (TCR). CAR-positive (+) T cells are engineered to express an extracellular single-chain variable fragment (scFv) with specificity for a particular tumor antigen linked to an intracellular signaling moiety that includes at least one costimulatory domain and at least one activation domain. CAR scFvs can be designed to target, for example, CD19, a transmembrane protein expressed by all normal B cells and cells in the B cell lineage, including B cell malignancies, including but not limited to NHL, CLL, and non-T cell ALL. In some embodiments, CARs are engineered such that the costimulatory domain is expressed as a separate polypeptide chain. Exemplary CAR T cell therapies and constructs are described in US Patent Publication Nos. 2013 / 0287748, 2014 / 0227237, 2014 / 0099309, and 2014 / 0050708, which are incorporated herein by reference in their entirety. In some embodiments, the immunotherapy is an autologous stem cell therapy, which can be performed according to methods described in the art, including those in the Examples. Any of the components of the CD19 CAR can include modifications and / or mutations that alter the properties of the CAR-T construct and / or the cells expressing the construct, such as modifications and / or mutations that affect tumor recognition, T cell cytokine production, T cell proliferation, T cell activation, T cell replication, T cell exhaustion, and T cell survival. Some examples of CAR constructs targeting CD19 are described in US Patent Publication No. 20170281766. In one embodiment, the CD19 CAR is a construct expressed in axicabtagene ciloleucel (Yescarta™). In one embodiment, the CD19 CAR is a construct expressed in Kymriah™. Additional CD19-directed CAR therapeutics that can be used in the methods of the present disclosure include, but are not limited to, JCAR017, JCAR015, JCAR014, Uppsala University anti-CD19 CAR (NCT02132624), and UCART19 (Cellectis).See Sadelain et al. Nature Rev. Cancer Vol. 3 (2003), Ruella et al., Curr Hematol Malig Rep., Springer, NY (2016), and Sadelain et al. Cancer Discovery (Apr 2013).

[0053] In one embodiment, to prepare a CD19-directed genetically modified autologous T cell immunotherapeutic, the patient's own T cells can be harvested and genetically modified ex vivo by retroviral transduction (e.g., gamma retroviral transduction) to express a chimeric antigen receptor (CAR) comprising a murine anti-CD19 single chain variable fragment (scFv) linked to the costimulatory domains of CD28 and CD3 zeta. In some embodiments, the CAR comprises a murine anti-CD19 single chain variable fragment (scFv) linked to the costimulatory domains of 4-1BB and CD3 zeta. The anti-CD19 CAR T cells can be expanded and infused back into the patient, where they can recognize and eliminate target cells expressing CD19. In some embodiments, the anti-CD19 CAR T cell therapy includes therapy with YESCARTA™ (Axi-cel™, axicabtagene ciloleucel), an example of one such CD19-directed genetically modified autologous T cell immunotherapeutic. See Kochenderfer, et al., (J Immunother 2009;32:689 702).

[0054] The T cells can be administered in a therapeutically effective amount. For example, a therapeutically effective amount of T cells is at least about 10 4 cells, at least about 10 5 cells, at least about 10 6 cells, at least about 10 7 cells, at least about 10 8 cells, at least about 10 9 pcs cells, or at least about 10 10 In another embodiment, the therapeutically effective amount of T cells can be about 104 cells, approximately 10 5 cells, approximately 10 6 cells, approximately 10 7 cells, or approximately 10 8 In some embodiments, the therapeutically effective amount of CAR T cells is about 2×10 6 cells / kg, approximately 3×10 6 cells / kg, approximately 4×10 6 cells / kg, approximately 5×10 6 cells / kg, approximately 6×10 6 cells / kg, approximately 7×10 6 cells / kg, approximately 8×10 6 cells / kg, approximately 9×10 6 cells / kg, approximately 1×10 7 cells / kg, approximately 2×10 7 cells / kg, approximately 3×10 7 cells / kg, approximately 4×10 7 cells / kg, approximately 5×10 7 cells / kg, approximately 6×10 7 cells / kg, approximately 7×10 7 cells / kg, approximately 8×10 7 cells / kg, or approximately 9 x 10 7 In some embodiments, the therapeutically effective amount of viable CAR-positive T cells is about 1×10 cells / kg of body weight. 6 Approximately 2 x 10 pieces 6 Between 1 x 10 and 2 x 10 CAR-positive viable T cells 8 In some embodiments, the therapeutically effective amount of CAR-positive viable T cells is about 1 x 10 per kg of body weight. 6 Pieces or about 2 x 10 6 Approximately 1 × 10 CAR-positive viable T cells 8 The same doses without the term about are also within the scope of the present disclosure.

[0055] In some embodiments, a therapeutically effective amount of viable CAR-positive T cells is about 0.4×10 8 From 2 x 10 8In some embodiments, the therapeutically effective amount of CAR-positive viable T cells is between about 0.4×10 8 pieces, approximately 0.5×10 8 pieces, approximately 0.6×10 8 pieces, approximately 0.7×10 8 pieces, approximately 0.8×10 8 pieces, approximately 0.9×10 8 pieces, approximately 1.0×10 8 pieces, approximately 1.1×10 8 pieces, approximately 1.2×10 8 pieces, approximately 1.3×10 8 pieces, approximately 1.4×10 8 pieces, approximately 1.5×10 8 pieces, approximately 1.6×10 8 pieces, approximately 1.7×10 8 pieces, approximately 1.8×10 8 pieces, approximately 1.9×10 8 Pieces or about 2.0 x 10 8 The term "viable T cells" has its conventional meaning in the art.

[0056] In some embodiments, the T cell composition comprises a pharma- ceutically acceptable carrier, diluent, solubilizer, emulsifier, preservative, and / or adjuvant. In some embodiments, the composition comprises an excipient. The components of the T cell composition are readily determined by one of skill in the art.

[0057] Methods for predicting response to CD19 CAR-T therapy The data presented herein support that the baseline sum of product diameters (SPD) of index lesions in a subject correlates with the degree of positive response to CD19 CAR-T therapy. Thus, in one embodiment, the present disclosure provides a method of predicting response to CD19 CAR-T therapy in a subject with cancer, comprising measuring the baseline sum of product diameters (SPD) of index lesions in the subject and determining an SPD range, where a low SPD range indicates a likelihood of a positive response to CAR-T therapy.

[0058] In one embodiment, certain values ​​of SPD can be useful in indicating the likelihood of response to CAR-T therapy. In one embodiment, the SPD can fall into one of at least four ranges or quartiles, with the lower range in which the SPD falls correlating with a higher likelihood of response to CD19 CAR-T therapy. In one embodiment, the four ranges include: SPD quartile 1, approximately 100 mm 2 (Including) ~ approx. 2000mm 2 (including), approx. 840mm 2 Median SPD, SPD quartile 2, approximately 2000 mm 2 (excluding) ~ approx. 3700mm 2 (including), approx. 2820mm 2 Median SPD, SPD quartile 3, approximately 3700 mm 2 (excluding) ~ approx. 6700mm 2 (including), approx. 5100mm 2 The median SPD and the quartile 4 SPD are approximately 6700 mm 2 (not included) ~ approx. 24000mm 2 (including), approx. 9300mm 2 Median SPD of 100 mg / kg / day. In one embodiment, baseline SPD is measured by site adjudication using the Cheson 2007 criteria (Neelapu SS and Locke FL, et al. Blood. 2016;128:LBA-6).

[0059] Clinical response may be measured by any parameter known in the art of cancer treatment, including CR, i.e. complete response, ORR, i.e. objective response rate, PR, i.e. partial response, and ORR, i.e. sustained response rate. In one embodiment, response is measured as sustained response rate assessed over at least one year of CD19 CAR-T treatment.

[0060] In one embodiment, a subject predicted to respond to CD-19 CAR-T therapy according to the present application is subsequently treated with a CD19 CAR-T therapy, where the subject has received at least one prior therapy that is not a CAR-T therapy, and the subject's SPD value is one of the SPD quartiles listed above. In one embodiment, if the subject is in SPD quartile 1, the subject is further or subsequently treated with a CD19 CAR-T therapy. In one embodiment, if the subject is in SPD quartile 2, the subject is further or subsequently treated with a CD19 CAR-T therapy. In one embodiment, the response is a sustained response to one year of therapy, the cancer is relapsed / refractory large diffuse B-cell lymphoma, and the treatment is ASCT of Yescarta as described in the ZUMA study (see Examples).

[0061] Exemplary Cancers "Cancer" refers to a broad group of various diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and growth can lead to the formation of malignant tumors that invade adjacent tissues and may metastasize to distant parts of the body via the lymphatic system or bloodstream. "Cancer" or "cancerous tissue" can include tumors. Examples of cancers that can be treated by the methods of the present disclosure include, but are not limited to, cancers of the immune system, including lymphomas, leukemias, myelomas, and other white blood cell malignancies. In some embodiments, the methods of the present disclosure are useful in treating cancers of the following types: bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal region cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, multiple myeloma, Hodgkin's disease, non-Hodgkin's lymphoma (NHL), primary mediastinal large B-cell lymphoma (PMBC), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), transformed follicular lymphoma, splenic marginal zone lymphoma (SMZL), esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic The present invention may be used to reduce tumor size in tumors derived from leukemia or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia (ALL) (including non-T-cell ALL), chronic lymphocytic leukemia (CLL), childhood solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers including asbestos-induced cancers, other B-cell malignancies, and combinations of the above cancers. In one particular embodiment, the cancer is multiple myeloma. In one embodiment, the method of the present application is suitable for cancers that respond to chemotherapy or radiation therapy, cancers that may be resistant to chemotherapy or radiation therapy, or refractory or relapsing cancers. In one embodiment, refractory cancer refers to a cancer that is not amenable to surgical intervention, the cancer does not respond initially to chemotherapy or radiation therapy, or the cancer becomes unresponsive over time.

[0062] In another embodiment, refractory (resistant) disease is indicated by less than a 50 percent decrease in lesion size with treatment in the absence of the development of new lesions. In contrast, disease progression is usually manifested as the appearance of new lesions, a 50% increase in the longest diameter of a previously identified lesion, or new / recurrent lesions in the bone marrow. In some embodiments, relapsing disease refers to the appearance of any new lesions after an initial complete remission has been achieved. In some embodiments, refractory or progressive disease is identified during response assessment after treatment. Relapses are usually symptomatic and are rarely identified based solely on routine imaging. Progression or relapse is usually characterized by a complete remission of the disease. It may present with the development of constitutional B symptoms (ie, fever, night sweats, weight loss), cytopenias, extranodal masses, or as symptomatic or asymptomatic enlargement of the lymph nodes, liver, or spleen.

[0063] In some embodiments, the cancer is acute lymphoblastic leukemia (ALL) (including non-T cell ALL), acute myeloid leukemia (AML), B cell prolymphocytic leukemia, B cell acute lymphoblastic leukemia (BALL), blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, chronic lymphocytic leukemia (CLL), chronic myelomatous leukemia (CML), chronic myelogenous leukemia, chronic or acute leukemia, diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), hairy cell leukemia, Hodgkin's disease, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma, myelodysplasia and myelodysplastic syndromes, non-Hodgkin's lymphoma (NHL). , plasma cell proliferative disorders (including indolent myeloma (smoldering multiple myeloma or indolent myeloma)), plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, plasmacytoma (including dysplasmocytosis, solitary myeloma, solitary plasmacytoma, extramedullary plasmacytoma and multiple plasmacytoma), POEMS syndrome (also known as Crow-Fukase syndrome, Takatsuki disease and PEP syndrome), primary mediastinal large B-cell lymphoma (PMBC), small cell or large cell follicular lymphoma, splenic marginal zone lymphoma (SMZL), systemic amyloid light chain amyloidosis, T-cell acute lymphoblastic leukemia (TALL), T-cell lymphoma, transformed follicular lymphoma or Waldenstrom's macroglobulinemia, or a combination thereof.

[0064] In some embodiments, the cancer is a hematological cancer. In one embodiment, the cancer is DLBCL, i.e. diffuse large B-cell lymphoma, PMBCL, i.e. primary mediastinal B-cell lymphoma, or TFL, i.e. transformed follicular lymphoma. In one embodiment, the cancer is relapsed or refractory diffuse large B-cell lymphoma (DLBCL). In one aspect, the invention provides a method of treating relapsed or refractory diffuse large B-cell lymphoma (DLBCL) not otherwise specified after two or more lines of systemic therapy in a patient, primary mediastinal large B-cell lymphoma, high-grade B-cell lymphoma, or DLBCL resulting from follicular lymphoma, by intravenous infusion of a suspension of CD19-directed genetically modified autologous T cells at about 1×10 per kg of body weight in a patient in need of treatment. 6 Approximately 2 x 10 pieces 6 Between 1 x 10 and 2 x 10 CAR-positive viable T cells 8 In one embodiment, the method comprises administering to the patient a dose of up to a maximal dose of viable CAR-positive T cells.

[0065] Methods of Treating Subpopulations of Cancer Subjects In some embodiments, the disclosed methods can be used to treat cancer in a subject, reduce tumor size, kill tumor cells, prevent tumor cell proliferation, prevent tumor growth, eliminate tumors from a patient, prevent tumor relapse, prevent tumor metastasis, induce remission in a patient, or any combination thereof. In certain embodiments, the methods induce a complete response. In other embodiments, the methods induce a partial response.

[0066] The data presented herein indicates that the baseline SPD, i.e., sum of products of diameters, of index lesions in a subject correlates with the degree of positive response to CD19 CAR-T therapy. Thus, in another embodiment, the present disclosure provides a method of treating cancer in a subject in need of treatment, comprising administering a therapeutically effective amount of a CD19 CAR-T therapy to a subject having a baseline SPD value in the first quartile of SPD. In one embodiment, the present disclosure provides a method of treating cancer in a subject in need of treatment, comprising administering a therapeutically effective amount of a CD19 CAR-T therapy to a subject having a baseline SPD value in the second quartile of SPD. The cancer may be any one of the cancers listed above. CD19 CAR-T therapy The therapeutic agent may be any one of the CD19 CAR-T therapeutic agents listed above. In one embodiment, baseline SPD is measured by any of the methods described above.

[0067] The higher the SPD quartile, the lower the response. Response and therapeutic benefit are correlated. In some embodiments, a complete response indicates therapeutic benefit. In some embodiments, a partial response indicates therapeutic benefit. In some embodiments, stable disease indicates therapeutic benefit. In some embodiments, an increase in overall survival indicates therapeutic benefit. In some embodiments, therapeutic benefit may constitute an improvement in time to disease progression and / or an improvement in symptoms or quality of life. In other embodiments, therapeutic benefit cannot be translated into an increase in duration of disease suppression, but rather a significant reduction in symptom burden resulting in an improvement in quality of life.

[0068] In another embodiment, the disclosure provides a method of treating cancer in a subject in need of treatment, comprising administering a therapeutically effective amount of a CD19 CAR-T therapy to a subject who has had 1-2, 3, 4, or 5 or more lines of prior treatment. In one embodiment, the disclosure provides a method of treating cancer in a subject in need of treatment, comprising administering a therapeutically effective amount of a CD19 CAR-T therapy to a subject who has had 1-2 lines of prior treatment. The cancer can be any one of the cancers listed above. CD19 The CAR-T therapy can be any one of the CD19 CAR-T therapies listed above. In some embodiments, the CD19 CAR-T therapy is used as a first line treatment.

[0069] The prior line of treatment can be any prior anti-cancer therapy, including, but not limited to, Bruton's tyrosine kinase inhibitors (BTKi), checkpoint inhibitors (e.g., anti-PD1 antibodies, pembrolizumab (Keytruda), cemiplimab (Libtayo), nivolumab (Opdivo), anti-PD-L1 antibodies, atezolizumab (Tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), anti-CTLA-4 antibodies, ipilimumab (Yervoy), anti-CD19 antibodies (e.g., blinatumomab), anti-CD52 antibodies (e.g., alemtuzumab), allogeneic stem cell transplant, anti-CD20 antibodies (e.g., rituximab), systemic chemotherapy with or without rituximab, rituximab, anthracyclines, xxxxxxxxx. Prior treatments can also include the CD19 CAR inhibitors of the present disclosure. It may be used in combination with T therapy. In one embodiment, eligible patients may have disease refractory to most current therapies or may relapse within one year after autologous hematopoietic stem cell transplantation (HSCT / ASCT). In some embodiments, the refractory disease is refractory large B-cell lymphoma (diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, transformed follicular lymphoma) or relapsed / refractory CLL. In another embodiment, eligible patients have relapsed or refractory LBCL with two or more prior systemic therapies. In another embodiment, the patient is suffering from acute lymphoblastic leukemia (ALL).

[0070] In one embodiment, the present disclosure provides that T cell immunotherapy with anti-CD19 CAR-T can induce a high proportion of durable responses with a manageable safety profile for patients aged 65 years or older. In another embodiment, the present disclosure provides that T cell immunotherapy with anti-CD19 CAR-T can induce a high proportion of durable responses with a manageable safety profile for patients aged under 65 years. In another embodiment, the present disclosure provides that anti-CD19 CAR-T therapy with axi-cel / Yescarta can induce a high proportion of durable responses with a manageable safety profile for patients aged 65 years or older. In another embodiment, the present disclosure provides that anti-CD19 CAR-T therapy with axi-cel / Yescarta can induce a high proportion of durable responses with a manageable safety profile for patients aged under 65 years.

[0071] In one embodiment, the present disclosure is a method of treating cancer in a subject in need of treatment. In one embodiment, the disclosure provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective CD19 CAR T therapy to a subject under 65 years of age. In one embodiment, the disclosure provides a method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective CD19 CAR T therapy to a subject under 65 years of age. In one embodiment, "CD19 CAR T therapy" refers to the number of CAR T cells administered. In one embodiment, "CD19 CAR T therapy" refers to a dosing regimen including the amount of CAR T cells with or without preconditioning and the number and timing of administrations.

[0072] The amounts of CAR T cells, dosing regimens, methods of administration, subjects, and cancers included within these methods are described elsewhere in this disclosure, either alone or in combination with another chemotherapeutic agent, with or without preconditioning, for any of the patients described elsewhere herein.

[0073] In one embodiment, the cancer is any of the cancers described above. In one embodiment, the CD19 CAR-T therapy is any of the CD19 CAR-T therapies described above. In one embodiment, the CD19 CAR-T therapy comprises a Yescarta therapy. In one embodiment, the CD19 CAR-T therapy comprises a therapy or protocol described in the ZUMA-1 trial (see Examples). In one embodiment, the response is a sustained response at one year of treatment, the cancer is relapsed / refractory large diffuse B-cell lymphoma, and the treatment is a Yescarta™ ASCT described in the ZUMA trial (see Examples).

[0074] In one embodiment, the CD19 CAR-T therapy is any of the CD19 CAR-T therapies described above. In one embodiment, the CD19 CAR-T therapy includes a Yescarta therapy. In one embodiment, the CD19 CAR-T therapy includes a treatment or protocol described in the ZUMA-1 trial (see Examples). In one embodiment, the response is a sustained response over one year of treatment, the cancer is relapsed / refractory large diffuse B-cell lymphoma, and the treatment is a Yescarta™ ASCT as described in the ZUMA-1 trial (see Examples).

[0075] In one embodiment, the cancer is refractory DLBCL, i.e., diffuse large B-cell lymphoma (DLBCL), primary mediastinal B-cell lymphoma (PMBCL) or transformed follicular lymphoma (TFL), optionally also having (i) failure to respond to last chemotherapy or relapsed within 12 months after ASCT, and (ii) receiving 500 mg / m 2 of cyclophosphamide and 30 mg / m 2 of fludarabine for 3 days, followed by 2 × 10 6 Axi-cel will be administered at CAR+ cells / kg.

[0076] In some embodiments, the method further comprises administering a chemotherapeutic agent or an additional therapeutic agent. In some embodiments, the chemotherapeutic agent selected is a lymphodepleting ) (preconditioning) chemotherapy agents. Beneficial preconditioning treatment regimens, along with correlative beneficial biomarkers, are described in U.S. Provisional Patent Applications 62 / 262,143 and 62 / 167,750, which are incorporated herein by reference in their entireties. These include, for example, the indicated beneficial doses of cyclophosphamide (200 mg / m 2 / day~2000mg / m 2 / day) and the specified dose of fludarabine (20 mg / m 2 / day~900mg / m 2 The present invention describes a method of conditioning a patient in need of T cell therapy comprising administering to the patient about 500 mg / m2 of engineered T cells per day. One such dosing regimen comprises administering to the patient about 500 mg / m2 of engineered T cells per day prior to administration of a therapeutically effective amount of the engineered T cells to the patient. 2 / day cyclophosphamide and approximately 60 mg / m 2 The present invention includes treating a patient comprising administering fludarabine to the patient daily for three days. Other examples of preconditioning regimens can be found, for example, in U.S. Patent No. 9,855,298.

[0077] A variety of additional therapeutic agents can be used in combination with the compositions described herein. For example, potentially useful additional therapeutic agents include PD-1 inhibitors such as nivolumab (Opdivo™), pembrolizumab (Keytruda™), pembrolizumab, pidilizumab (CureTech), and atezolizumab (Roche). Additional therapeutic agents suitable for use in combination with the compositions and methods described include, but are not limited to, ibrutinib (Imbruvica™), ofatumumab (Arzera™), rituximab (Rituxan™), bevacizumab (Avastin™), trastuzumab (Herceptin™), trastuzumab emtansine (Kadcyla™), imatinib (Gleevec™), cetuximab (Erbitux™), panitumumab (Vectibix™), catumaxomab, ibritumomab, ofatumuma ... Atumumab, tositumomab, brentuximab, alemtuzumab, gemtuzumab, erlotinib, gefitinib, vandetanib, afatinib, lapatinib, neratinib, axitinib, masitinib, pazopanib, sunitinib, sorafenib, toceranib, lestaurtinib, axitinib, cediranib, lenvatinib, nintedanib, pazopanib, regorafenib, semaxanib, sorafenib, sunitinib, tivozanib, toceranib, vandetanib, entrectinib, cabozantinib, imatinib, dasatinib, nilotinib, ponatinib mTOR inhibitors such as rivaroxaban, radotinib, bosutinib, lestaurtinib, ruxolitinib, pacritinib, cobimetinib, selumetinib, trametinib, binimetinib, alectinib, ceritinib, crizotinib, aflibercept, adipotide, denileukin diftitox, everolimus and temsirolimus; hedgehog inhibitors such as sonidegib and vismodegib; CDK inhibitors such as CDK inhibitor (palbociclib); Bruton's tyrosine kinase inhibitors (BTKi); checkpoint inhibitors (e.g., anti-PD1 antibodies, pembrolizumab, These include izumab (Keytruda), cemiplimab (Libtayo), nivolumab (Opdivo), anti-PD-L1 antibodies, atezolizumab (Tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), anti-CTLA-4 antibodies, ipilimumab (Yervoy), anti-CD19 antibodies (e.g., blinatumomab), anti-CD52 antibodies (e.g., alemtuzumab), allogeneic stem cell transplant, anti-CD20 antibodies (e.g., rituximab), systemic chemotherapy with or without rituximab, rituximab, anthracyclines, cytokines, other anti-inflammatory agents, and the like.Pretreatment may also be used in combination with the CD19 CAR T therapy of the present disclosure.

[0078] Examples of other chemotherapeutic agents that may be used with the anti-CD19 therapy of the present disclosure include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN™); alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphaoramide, and trimethylolmelamine. ethylenimines and methylmelamines, including the ethylenimines and methylmelamines regimes; chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichine, phenesterine, prednimustine, trofosfamide, nitrogen mustards such as uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; aclacinomysins, aclacinomycin, Cutinomycin, autramycin, azaserine, bleomycin, Cutinomycin, calicheamicin, carabicin, carminomycin, Rudinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin Bicine, esorubicin, idarubicin, marcellomycin n), mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin isin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, th Antibiotics such as vermicidin, ubenimex, zinostatin, and zorubicin; Antimetabolites such as thotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, etc. pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU, etc.; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, etc.; anti-adrenal agents such as aminoglutethimide, mitotane, trilostane, etc.; folic acid replacement fluids such as frolinic acid, etc.; aceglatone; Aldophosphamide glycosides; Aminolevulinic acid; Amsacrine; Bestrabucil; Bisantrene; Edatraxate; Defofamine; Demecolcine; Diazicon; Elformithine; Elformithine acetate putinium; etoglucide; gallium nitrate; hydroxurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK (trademark); razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2''-trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, such as paclitaxel (TAXOL (trademark), Bristol-Myers Squibb) and docetaxel (TAXOTERE™, Rhone-Poulenc Rorer); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronic acid; CPT-11; the topoisomerase inhibitor RFS2000; difluoromethylomitin (DMFO); retinoic acid derivatives such as Targretin™ (bexarotene), Panretin™ (alitretinoin); ONTAK™ (denileukin diftitox); esperamicin; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above. In some embodiments, compositions comprising immune effector cells expressing a CAR and / or TCR disclosed herein are administered in combination with anti-hormonal agents that act to regulate or inhibit the action of hormones on tumors, such as, for example, tamoxifen, raloxifene, aromatase-inhibiting 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, Antiestrogens, including onapristone, and toremifene (Fareston); and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharma- ceutical acceptable salts, acids, or derivatives of any of the above. Also, where appropriate, combinations of chemotherapy agents, including but not limited to CHOP, i.e., cyclophosphamide (Cytoxan™), doxorubicin (hydroxydoxorubicin), vincristine (Oncovin™), and prednisone, are administered.

[0079] In some embodiments, the antigen binding molecules (e.g., antibodies listed above), transduced (or otherwise engineered) cells (e.g., CARs), and chemotherapeutic agents are each administered in an amount effective to treat the subject's disease or condition, either alone or in combination with other agents and therapeutic agents described herein.

[0080] How to predict the likelihood of a relapse The data presented herein indicates that the number of prior lines of therapy in a subject correlates with the degree or likelihood of relapse after CD19 CAR-T therapy. Thus, in one embodiment, the present disclosure provides a method for predicting the likelihood of relapse after CD19 CAR-T therapy in a subject with cancer, comprising determining the number of prior lines of therapy in the subject and determining which of four ranges the number falls within, where the higher the number of prior lines of therapy, the higher the likelihood of relapse after therapy for the subject is predicted to be.

[0081] In one embodiment, the four ranges of number of prior lines of therapy are 1-2, 3, 4, and 5 or more. In one embodiment, the prior lines of therapy include any one or more of a preceding anti-CD20 monoclonal antibody, an anthracycline, an appropriate standard of care, and combinations thereof. In one embodiment, the prediction method is for determining whether a subject with any of the above ranges of prior lines of therapy has CD19 It further comprises administering a CAR-T therapy. In one embodiment, the therapy is administered to a subject who has received 1-2 lines of prior therapy. In one embodiment, the extent of relapse is measured within 12 months of treatment, the cancer is relapsed / refractory large diffuse B-cell lymphoma, and the therapy is ASCT of Yescarta as described in the ZUMA trial (see Examples).

[0082] Methods for predicting likelihood of response to CAR-T therapy in a subject with cancer The data presented herein also indicates that the number of prior lines of therapy in a subject correlates with the degree of positive response to CD19 CAR-T therapy.Accordingly, the present disclosure also provides a method for predicting the likelihood of response to CD19 CAR-T therapy in a subject with cancer, comprising measuring the number of prior lines of therapy in the subject at baseline and determining which of four ranges the number falls into, wherein the lower the range the subject falls into, the higher the likelihood of sustained response to CD19 CAR-T therapy.

[0083] In one embodiment, the four ranges for the number of prior lines of therapy are 1-2, 3, 4, and greater than 5. In one embodiment, the prior lines of therapy include a prior anti-CD20 monoclonal antibody, anthracycline, cyclophosphamide, fludarabine, any one or more of the appropriate standard therapies, and combinations thereof.

[0084] In one embodiment, the cancer is any of the cancers listed above. In one embodiment, the CD19 CAR-T therapy includes any of the therapies disclosed above.

[0085] In one embodiment, the predictive method further comprises administering a CD19 CAR-T therapy to a subject with any of the above ranges of lines of prior therapy. In one embodiment, the therapy is administered to a subject who has received 1-2 lines of prior therapy. In one embodiment, the response is a sustained response over one year of therapy, the cancer is relapsed / refractory large diffuse B-cell lymphoma, and the treatment is ASCT of Yescarta as described in the ZUMA trial (see Examples).

[0086] Clinical response may be measured by any parameter known in the art of cancer treatment, including CR, i.e. complete response, ORR, i.e. objective response rate, PR, i.e. partial response, and ORR, i.e. sustained response rate. In one embodiment, response is measured as sustained response assessed over one year of CD19 CAR-T treatment.

[0087] The following non-limiting examples and data illustrate various aspects and features associated with the disclosed cell and therapeutic methods and uses. In some embodiments, the methods and uses of the compounds include , resulting in surprising, unexpected and unexpected results and data. Although the utility of the disclosed methods is illustrated through the use of several cells or compounds that can be used together, one of skill in the art will appreciate that equivalent results can be obtained using a variety of other cells or compounds that fall within the scope of the present disclosure.

[0088] In the context of these examples, ASCT means autologous stem cell transplant, DLBCL means diffuse large B-cell lymphoma, ECOG means Eastern Cooperative Oncology Group performance status, IPI means International Prognostic Index, PD means progressive disease, PMBCL means primary mediastinal B-cell lymphoma, SPD means sum of products of diameters, and TFL means transformed follicular lymphoma. EXAMPLES

[0089] Example 1 Outcomes by prior line of treatment in a pivotal phase 2 study of axicabtagene-ciloleucel in patients with refractory large B-cell lymphoma Axi-cbutagen-ciloleucel (axi-cel) is an autologous chimeric antigen receptor (CAR) T-cell therapy that recognizes and eliminates CD19-expressing cells. In a pivotal phase 1 / 2 multicenter trial (ZUMA-1), 108 patients with refractory large B-cell lymphoma were treated (median follow-up, 15.4 months) and showed an 82% objective response rate, including 58% complete response (CR), 42% durable response, including 40% for CR, 13% grade ≥3 cytokine release syndrome (CRS), and 28% grade ≥3 neurological events.

[0090] In phase 1, subjects (n=7) had refractory DLBCL, i.e., diffuse large B-cell lymphoma (DLBCL), primary mediastinal B-cell lymphoma (PMBCL), or transformed follicular lymphoma (TFL). Phase 2 consisted of two cohorts. Subjects in cohort 1 (n=77) had refractory DLBCL. Subjects in cohort 2 (n=24) had refractory PMBCL / TFL. Additional eligibility criteria included (i) failure to respond to last chemotherapy or relapse within 12 months after ASCT and (ii) prior anti-CD20 monoclonal antibody and anthracycline. All subjects received 500 mg / m 2 of cyclophosphamide and 30 mg / m 2 All patients received a conditioning regimen of 2 × 10 6 Patients were administered Axi-cel at CAR+ cells / kg. 99% of enrolled subjects were successfully manufactured and 91% of enrolled subjects were administered.

[0091] Treatment included several steps in the following order: screening, leukapheresis, and conditioning chemotherapy followed by Axi-cel infusion. Manufacturing took place between leukapheresis and conditioning chemotherapy. Axi-cel infusion took place on day 0. Post-treatment evaluations and long-term follow-up took place from day 7 onwards during the follow-up period. The first tumor evaluation took place on day 28. Bridging chemotherapy was not allowed by the study protocol.

[0092] The following parameters were used for evaluation and statistical analysis: safety and efficacy outcomes were evaluated by number of prior lines of therapy: 1-2, 3, 4 or ≥5 prior lines; autologous stem cell transplant was considered as a prior line of therapy; safety and efficacy outcomes were evaluated by quartiles of tumor burden; tumor burden was estimated as the sum of products of diameters (SPD) of the index lesion by institutional adjudication according to the Cheson 2007 criteria (Neelapu SS and Locke FL, et al. Blood. 2016;128:LBA-6); and / or the SPD of the index lesion may not be representative of the entirety of the patient's disease.

[0093] These results showed that patients with more lines of prior therapy were not only more likely to relapse after ASCT but also had higher International Prognostic Index scores and disease stages, which is consistent with a higher SPD (Table 1).

[0094] [Table 1]

[0095] Table 1: Baseline characteristics. ASCT, autologous stem cell transplant, DLBCL, diffuse large B-cell lymphoma, ECOG, Eastern Cooperative Oncology Group performance status, IPI, International Prognostic Index, PMBCL, primary mediastinal B-cell lymphoma, SPD, sum of products of diameters, TFL, transformed follicular lymphoma.

[0096] Additionally, product characteristics were similar across prior lines of therapy (see Tables 2 and 3). Lines of therapy were enumerated prior to enrollment in ZUMA-1.

[0097] [Table 2]

[0098] Table 2: Product characteristics. a Doubling time was calculated by the following calculation: doubling time = (ln[2] × duration) / (ln[total viable cells at harvest / total viable cells on day 3]). b Phenotypic analysis of anti-CD19 CAR T cells by flow cytometry included the surface markers CD3, CD4, CD8, CCR7, and CD45RA. CCR7 and CD45RA were used to identify TN (CCR7 + CD45RA + ) T cells and TCM (CCR7 + CD45RA - ) T cells and TEM (CCR7 - CD45RA - ) T cells and TEFF (CCR7 - CD45RA + ) T cells were distinguished. CR is complete response, ORR is objective response rate, PR is partial response, TCM is central memory T cells, TEFF is effector T cells, TEM is effector memory T cells, and TN is naive T cells.

[0099] [Table 3]

[0100] Table 3: Response rates by prior line of treatment. CR is complete response, ORR is objective response rate, and PR is partial response.

[0101] Furthermore, the rate of grade ≥3 CRS was similar across lines of prior therapy (data not shown).For patients with ≥5 prior lines of therapy, there was a trend towards increased rates of grade ≥3 serious adverse events and neurological events.

[0102] Table 4 shows the expansion of CAR T cells by prior line of therapy. Counts were taken prior to enrollment in ZUMA-1.

[0103] [Table 4]

[0104] Table 4: CAR T cell expansion by prior line of treatment. AUC 0-28 is the cumulative level of CAR+ cells per µL of blood during the first 28 days after axi-cel, where axi-cel is axocabtagene ciloleucel and CAR is a chimeric antigen receptor.

[0105] Baseline characteristics suggest that patients with more lines of prior therapy had higher IPI and disease stage. In addition, an evaluation of outcomes by indexed SPD was performed. The results, shown in Table 5, show that the lowest SPD quartile (median 840 mm 2 ) suggest that patients with substantial disease have

[0106] [Table 5]

[0107] Table 5: SPD by quartiles. Square brackets indicate inclusive values. Round brackets indicate exclusive values. b The SPD for one patient could not be obtained.

[0108] As SPD quartiles increased, the rate of durable response decreased (Figure 1). Grade ≥ 3 CRS and neurologic events occurred least frequently in patients in the lowest SPD quartile.

[0109] These results confirm that axi-cel demonstrated long-term clinical benefit for patients with refractory large B-cell lymphoma, regardless of number of prior lines of therapy and SPD. Axi-cel also produced similar product characteristics across all lines of therapy. CAR peak proliferation, area under the curve, and range were comparable across patients with 1-4 prior lines of therapy. Furthermore, the study demonstrated that higher one-year sustained response rates were observed in patients with lower SPD, and lower rates of CRS and neurological events were observed in patients with index lesion SPD in the lowest SPD quartile.

[0110] Example 2 Long-term activity of axicabtagene-ciloleucel in refractory large B-cell lymphoma In this study, over 100 patients with refractory large B-cell lymphoma were treated with axicabtagene ciloleucel and their cancer progression was followed for more than 2 years after a single treatment. These results indicate that axicabtagene ciloleucel can result in durable responses and a median overall survival of more than 2 years. Results were similar across all patient subgroups, including the majority of patients with activated B-cell-like lymphoma, double expressor lymphoma, and high-grade B-cell lymphoma.

[0111] One hundred and nineteen patients were enrolled and 108 were treated. Patient distribution across number of prior lines of therapy was as follows: Phase 1: median (IQR) 3 (3–4); 1 line (0 patients); 2 lines (1 patient, 14%), 3 lines (6 patients, 86%). Phase 2: median (IQR) 3 (2–4); 1 line (3 patients, 3%); 2 lines (28 patients, 28%), 3 lines (70 patients, 69%). Furthermore, in Phase 1, 1 patient (14%) had a history of primary refractory disease and 1 patient (14%) had a history of resistance to two consecutive lines. In Phase 2, 26 patients (26%) had a history of primary refractory disease and 54 patients (53%) had a history of resistance to two consecutive lines. Patients may have received other therapy after primary refractory disease.

[0112] Regarding cancer characterization, 52 (70%) of 74 patients evaluated for cellular origin had germinal center B cell-like disease and 18 (24%) had activated B cell-like disease. Of 47 patients with pretreatment tumor samples, 30 (64%) had double expresser B cell lymphoma and 7 (15%) had high-grade B cell lymphoma, including 1 (2%) with triple-hit high-grade B lymphoma, 4 (9%) with double-hit high-grade B cell lymphoma, and 2 (4%) with unspecified high-grade B cell lymphoma.

[0113] One hundred and one patients evaluable for activity in phase 2 were followed for a median of 27.1 months (IQR 25.7-28.8). Of the 101 patients, 84 (83%) had an objective response to treatment by institutional assessment, 59 (58%) had a complete response, and 25 (25%) had a partial response. Ten (10%) patients had stable disease, five (5%) had progressive disease as their best response, and two (2%) were not evaluable.

[0114] Median response duration was 1 month (IQR 1-1). Eleven of 33 patients with a partial response at 1 month and 11 of 24 patients with stable disease at 1 month went on to achieve a complete response, with many switchovers occurring by 6 months. Of 33 patients with double expressor lymphoma and high-grade B-cell lymphoma, 30 (91%) had an objective response and 23 (70%) had a complete response. Median response duration for all 101 patients was 11.1 months (95% CI 4.2-not estimable).

[0115] Duration of response was consistent across key baseline and clinical covariates. Median progression-free survival was 5.9 months (95% CI 3.3 to 15). Estimated proportions of patients with progression-free survival at 24 months were 72% (95% CI 56.0 to 83.0) of patients with a complete response at 3 months, 75% (31.5 to 93.1) of patients with a partial response at 3 months, and 22.2% (3.4 to 51.3) of patients with stable disease at 3 months. Median overall survival was not reached (95% CI 12.8 to not estimable), and the estimated 24-month survival proportion was 50.5% (95% CI 40.2 to 59.7). No patients were lost to follow-up.

[0116] Sustained responses at 24 months were associated with higher peak CAR T cell concentrations and areas under the curve during the first 28 days after infusion. By 24 months, 11 of 32 evaluable patients (34%) maintained a sustained response and no longer had detectable engineered T cells. The safety profile 2 years after infusion was similar to that of the shorter timeline.

[0117] Analysis of progression-free survival by response at 3 months suggests that achieving a complete or partial response at 3 months may be predictive of long-term response durability (Figure 2). Of 35 evaluable patients with ongoing responses, 6 (17%) had detectable B cells in the peripheral blood. At 9 months, 20 (61%) of 33 evaluable patients had detectable B cells, and at 24 months, 24 (75%) of 32 evaluable patients had detectable B cells.

[0118] These results suggest that patients with refractory large B-cell lymphoma treated with a single infusion of axicabtagene-ciloleucel showed durable responses lasting longer than 2 years and did not require further consolidation therapy. In this population of patients refractory to several lines of therapy, including the majority of patients with activated B-cell-like lymphoma, double expressor lymphoma, and high-grade B-cell lymphoma, results were similar across all patient subpopulations. Median overall survival was not reached at 2 years, with an estimated 24-month survival rate of 50.5% (95% CI 40.2-59.7). In comparison, the expected median overall survival with conventional therapy is approximately 6 months, and the 2-year overall survival is approximately 20%.

[0119] Despite targeting CD19 and the expected induction of B-cell aplasia, the frequency of late grade 3 or worse serious infections was low. Seventy-five percent of evaluable patients with sustained responses showed evidence of B-cell recovery by 24 months, with some patients experiencing the onset of B-cell recovery at 9 months. The B-cell recovery in these sustained responders suggests that durable responses in adults with lymphoma may not require long-term persistence of functional CAR T cells.

[0120] Example 3 Efficacy and safety results in patients aged 65 years or older from a phase 1 / 2 study of axicabtagene-ciloleucel (Axi-Cel) in refractory large B-cell lymphoma (LBCL) Eligible patients with refractory large B-cell lymphoma (LBCL) underwent leukapheresis and received 500 mg / m 2 (intravenous) cyclophosphamide and 30 mg / m 2(intravenous) fludarabine was administered, where both were administered at a dose of 2 × 10 6 Axicabtagene ciloleucel was administered at a dose of 100 CAR-positive viable T cells / kg on days 5, 4, and 3 prior to axicabtagene ciloleucel (day 0). 108 patients were treated. Patients ≥65 years (n=27) vs <65 years (n=81) had a median age of 69 vs 55 years (y), 81% vs 63% were male, 70% vs 36% had IPI score 3-4, 59% vs 57% had ECOG 1, 81% vs 84% ​​were stage III / IV, 67% vs 72% had ≥3 prior therapies, and median (range) tumor burden by SPD was 3790 (600-16764) mm 2 3574(171~23297)mm 2 CAR T cell expansion to peak levels (43 cells / μl vs. 35 cells / μl) or area under the curve (562 days × cells / μl vs. 448 days × cells / μl) were similar in patients aged 65 years (y) or older vs. those younger than 65 years, respectively. With regard to disease type, 74% vs. 79% had DLBCL, 0% vs. 10% had PMBCL, and 26% vs. 11% had TFL. 19% vs. 30% had prior ASCT. The pre-enrollment refractory subgroup was categorized as follows: 4% vs. 2% were refractory to first-line therapy, 78% vs. 73% were refractory to second-line or subsequent line therapy, and 19% vs. 25% relapsed after ASCT.

[0121] The median follow-up was 27.1 months for phase 2 patients (n=101). Efficacy by age group was categorized as follows: ORR for patients ≥65 years (n=24) and <65 years (n=77) was 92% and 81%, respectively (CR rate 75% and 53%; PR 17% vs. 27%), with ongoing responses in 42% and 38% of patients (ongoing CR 42% and 35%). 24-month OS rates were 54% for patients ≥65 years and 49% for patients <65 years. 100% of patients ≥65 years and 65% of patients <65 years were 65% and 65% were 65% and 65%, respectively. Grade ≥3 AEs were observed in 98% of patients aged ≥65 years and ≥65 years. Grade ≥3 neurological events and cytokine release syndrome occurred in 44% vs. 28% and 7% vs. 12% of patients aged ≥65 years and ≥65 years, respectively. Rates of grade ≥3 cytokine release syndrome (CRS) and neurological events (NE) were similar across age groups. CRS included fever, hypotension, and hypoxia. NE included encephalopathy, confusion, aphasia, agitation, and delirium. Any grade and grade ≥3 cytopenias (cytopenia, thrombocytopenia, neutropenia, anemia) were consistent across age groups. 26% and 32% of patients ≥65 years and ≥65 years received intravenous immunoglobulin therapy, respectively.

[0122] Compared with patients younger than 65 years, patients aged 65 years or older showed disease progression as the best response to the last prior treatment, with an IPI score of 3 to 4 (age 60 years or older is a component of the IPI score). Axi-cel could induce a high proportion of durable responses with a manageable safety profile in patients aged 65 years or older and in patients younger than 65 years. Elderly patients with refractory LBCL generally have limited treatment options. Results of the 2-year analysis showed treatment efficacy (83% objective response rate, 58% complete response rate, 39% sustained response; N=101, median follow-up 27.1 months) and safety (late adverse events were mainly manageable infections, there were no late axi-cel-related occurrences of cytokine release syndrome, neurological events, or death; N=108).

Claims

1. 1. A method of predicting a response to CD19 CAR-T therapy in a subject having cancer, comprising measuring baseline SPD in said subject and determining an SPD range, wherein a low SPD range indicates a likelihood of a positive response to said CAR-T therapy.

2. 2. The method of claim 1, wherein the SPD can fall into one of four ranges, where the lower the range the SPD falls into, the higher the likelihood of response to CD19 CAR-T therapy.

3. The four ranges are as follows: SPD quartile 1, approximately 100 mm ２ (Including) ~ approx. 2000mm ２ (Including), median SPD of about 840; SPD quartile 2, approximately 2000 mm ２ (excluding) ~ approx. 3700 mm ２ (including) approx. 2820mm ２ Median SPD of SPD quartile 3, approximately 3700 mm ２ (excluding) ~ approx. 6700 mm ２ (Including) approx. 5100mm ２ The median SPD of SPD quartile 4, approximately 6700 mm ２ (Not included) ~ approx. 24,000 mm ２ (Including) approx. 9,300 mm ２ Median SPD of The method of claim 2 , comprising:

4. 4. The method of any one of claims 1-3, wherein if the baseline SPD value is in the SPD quartile 1-4, the subject is subsequently treated with a CD19 CAR-T therapy.

5. 1. A method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of a CD19 CAR-T therapy to a subject having a baseline SPD level in the SPD quartile 1-4.

6. 1. A method of predicting the likelihood of recurrence after CD19 CAR-T therapy in a subject having cancer, comprising determining the number of prior lines of therapy in the subject and determining into which one of four ranges the number falls, wherein the higher the number of prior lines of therapy, the higher the predicted likelihood of recurrence after CD19 CAR-T therapy for the subject.

7. 1. A method of predicting likelihood of sustained response to CD19 CAR-T therapy in a subject with cancer, comprising measuring the number of prior lines of therapy in the subject at baseline and determining which one of four ranges the number falls within, wherein a lower range indicates a greater likelihood of sustained response to CD19 CAR-T therapy.

8. The method according to claim 6 or 7, wherein the number of prior treatment lines is in the range of 1 to 2, 3, 4, or 5 or more.

9. The method of any one of claims 6 to 8, further comprising subsequently administering a CD19 CAR-T therapy to the subject who has had 1 to 2, 3, 4, or 5 or more lines of prior therapy.

10. 1. A method of treating cancer in a subject in need of treatment, comprising administering a therapeutically effective amount of a CD19 CAR-T therapy to a subject, wherein the subject has had 1 to 2, 3, 4, or 5 or more lines of prior therapy.

11. The method of any one of claims 1 to 10, wherein the cancer is a hematological cancer or relapsed / refractory diffuse large B-cell lymphoma.

12. 12. The method of any one of claims 1 to 11, wherein the CD19 CAR-T treatment comprises treatment with axicabtagene ciloleucel (Yescarta), tisagenlecleucel (Kymriah), JCAR017, JCAR015, JCAR014, Uppsala University anti-CD19 CAR (NCT02132624) or UCART19.

13. 1. A method of predicting long-term durability of response to treatment of cancer with an anti-CD19 CAR-T cell therapy in a patient in need of such treatment, comprising assessing progression free survival three months after a single administration of the therapeutic agent, wherein achievement of a complete or partial response at three months is predictive of long-term durability of response in said patient.

14. 14. The method of claim 13, wherein the anti-CD19 CAR-T treatment comprises treatment with axicabtagene ciloleucel (Yescarta), tisagenlecleucel (Kymriah), JCAR017, JCAR015, JCAR014, Uppsala University anti-CD19 CAR (NCT02132624) or UCART19.

15. 15. The method of claim 13 or 14, wherein the cancer is a hematological cancer.

16. 16. The method of claim 15, wherein the cancer is relapsed / refractory diffuse large B-cell lymphoma.

17. The method of any one of claims 13 to 16, wherein the long-term response durability comprises a complete or partial response lasting for more than 9 months, more than 12 months, more than 18 months, or more than 24 months.

18. The method of any one of claims 1 to 17, wherein the subject is 65 years of age or older.

19. The method of any one of claims 1 to 17, wherein the subject is under 65 years of age.

20. The method of any one of claims 1 to 19, wherein the CD19 CAR-T therapeutic is administered as a first line therapy.

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