WO1997045142A1 - Prevention de la reaction du greffon contre l'hote au moyen de lymphocytes thymo-dependants contenant des polynucleotides codant des marqueurs de selection negative - Google Patents

Prevention de la reaction du greffon contre l'hote au moyen de lymphocytes thymo-dependants contenant des polynucleotides codant des marqueurs de selection negative Download PDF

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WO1997045142A1
WO1997045142A1 PCT/US1997/009040 US9709040W WO9745142A1 WO 1997045142 A1 WO1997045142 A1 WO 1997045142A1 US 9709040 W US9709040 W US 9709040W WO 9745142 A1 WO9745142 A1 WO 9745142A1
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cells
host
graft
versus
administered
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PCT/US1997/009040
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English (en)
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Nikhil Munshi
David L. Ennist
William F. Jacob
Yawen L. Chiang
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Genetic Therapy, Inc.
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Priority to IL12721197A priority Critical patent/IL127211A0/xx
Priority to AU30796/97A priority patent/AU719930B2/en
Priority to JP09542889A priority patent/JP2000511539A/ja
Priority to EP97925747A priority patent/EP0977595A4/fr
Priority to NZ333053A priority patent/NZ333053A/en
Publication of WO1997045142A1 publication Critical patent/WO1997045142A1/fr
Priority to NO985522A priority patent/NO985522L/no

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/001—Preparations to induce tolerance to non-self, e.g. prior to transplantation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/52—Purines, e.g. adenine
    • A61K31/522—Purines, e.g. adenine having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/461—Cellular immunotherapy characterised by the cell type used
    • A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/4643—Vertebrate antigens
    • A61K39/4644—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/38—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the dose, timing or administration schedule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/46—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the cancer treated
    • A61K2239/48—Blood cells, e.g. leukemia or lymphoma

Definitions

  • This invention relates to the prevention of graft- versus-host disease, or GVHD, in patients who have received allogeneic bone marrow transplants. More particularly, this invention relates to the prevention of graft-versus-host disease in patients who are suffering from relapsing or persistent leukemia, subsequent to an allogeneic T-cell depleted bone marrow transplant, by administering to such patients T-cells which include a polynucleotide encoding a negative selective marker, followed by the administration of an interaction agent or prodrug to kill the T-cells, in particular, those T-cells which are graft-versus-host reactive, before graft-versus-host disease develops.
  • T-cells which include a polynucleotide encoding a negative selective marker
  • this invention relates to the prevention of graft-versus-host disease in connection with the treatment of diseases or disorders wherein the treatment includes bone marrow ablation followed by the administration of an allogeneic bone marrow transplant, such as a T-cell depleted bone marrow transplant.
  • an allogeneic bone marrow transplant such as a T-cell depleted bone marrow transplant.
  • T-cells which include a polynucleotide encoding a negative selective marker also are administered to a patient, and an interaction agent or prodrug is administered to the patient to kill the T- cells, in particular, those T-cells which are graft-versus- host reactive, before graft-versus-host disease develops.
  • Hematological malignancies such as, for example, multiple myeloma (MM) , chronic myelogenous leukemia (CML) , acute myeloid leukemia (AMD , and acute lymphoblastic leukemia (ALL) , affect thousands of Americans per year. For example, multiple myeloma affects approximately 4 in 100,000 Americans per year. There were 12,800 new cases in 1993. Multiple myeloma comprises slightly more than 1% of all types of malignancies and slightly more than 10% of all hematological malignancies. (Barlogie, et al., JAMA. Vol. 268, pgs. 2946-2951 (1992); Barlogie, et al. , Blood. Vol.
  • Allogeneic bone marrow transplantation has emerged as an effective treatment modality for selected patients with multiple myeloma (Bortin, et al., 1986). 268 patients have received allotransplants in different trials. (Barlogie, et al., Seminars in Hematolo ⁇ v, Vol. 32, pgs. 31-44 (1995)). Fifty percent of the patients died within one year, about 40 percent achieved a complete response, and the four year projected event - free survival is approximately 35 percent.
  • Allogeneic bone marrow transplantation is thought to be curative, in part, because of an anti-tumor (i.e., graft- versus-myeloma, or GVM) effect derived from the adoptive transfer of immunocompetent cells in the donor graft.
  • an anti-tumor i.e., graft- versus-myeloma, or GVM
  • GVM graft- versus-myeloma
  • T-cell depletion In order to decrease the early mortality from allogeneic bone marrow transplantation, T-cell depletion has been employed. T-cells may be depleted from the allograft by any of several techniques, including density gradient centrifugation, soybean lectin agglutination and E-rosette formation, centrifugal elutriation, cytotoxic drugs or corticosteroids, anti-T-cell monoclonal antibodies, and positive selection of CD34+ cells. (Champlin, Journal of Hematotherapy. Vol. 2, pgs. 27-42 (1993); Reisner, et al. , The Lancet. Vol. 2, pgs. 327-331 (1981); Waldmann, et al. , The Lancet. Vol.
  • This technique which consistently removes 2.5-3.0 log, 0 clonable T-cells, first employs differential agglutination with soybean lectin to remove mature leukocytes, including T- cells, B-cells, monocytes, and granulocytes, followed by E- rosette depletion for removal of residual T-cells.
  • This technique has been used to treat a population of over 200 HLA-matched related transplants given to leukemia patients. In this population, the incidence of Grade II acute GVHD has been 5%, and Grades III and IV acute GVHD have not been observed. (O'Reilly, et al., Bone Marrow Transplantation. Vol. 3, pgs. 3-6 (1988) .)
  • T-cell depletion decreases the incidence of GVHD
  • T-cell depletion also increases the risk of early relapse, incomplete immunological reconstitution, graft failure, and Epstein-Barr Virus-related lymphoma.
  • peripheral blood mononuclear cells can induce complete and long-term remission in such patients, such therapy may be associated with GVHD.
  • GVHD Bar, et al., J. Clin. Oncol.. Vol. 11, pg. 513 (1993); Drobyski, et al. , 1993; Slavin, et al. , 1990.
  • Allogeneic bone marrow transplantation following T-cell depletion also has been associated with increased incidence of bone marrow engraftment failure.
  • Epstein-Barr Virus (EBV) lymphoproliferative disorders.
  • EBV Epstein-Barr Virus
  • These lymphoproliferative disorders are often unresponsive to standard forms of therapy.
  • the Memorial Sloan-Kettering Bone Marrow Transplantation Group has reported on 7 patients developing lymphoproliferative disorder who were treated with donor lymphocyte infusion. (Papadopoulos, et al., Blood. Vol.
  • T- cells may be transduced with the Herpes Simplex Virus thymidine kinase (TK) gene ex vivo, and then be administered to a patient with hematopoietic stem cells. If the patient develops GVHD, ganciclovir may be administered to the patient in order to deplete the transduced T-cells.
  • TK Herpes Simplex Virus thymidine kinase
  • Such patients are given T-cells which have been genetically engineered to include a polynucleotide encoding a negative selective marker.
  • an interaction agent or prodrug is administered to the patient, whereby the genetically engineered T-cells are killed, and the development of GVHD is prevented.
  • the interaction agent may be administered to the patient at the time of development of GVHD, whereby the GVHD is treated through the killing of the transduced T-cells.
  • diseases or disorders include, but are not limited to, solid tumor malignancies, and acquired or genetic immunologic or hematopoietic diseases.
  • T-cells genetically engineered with a polynucleotide encoding a negative selective marker are administered to the patient. After the cells have remained in the patient for an amount of time sufficient to provide a therapeutic effect, an interaction agent or prodrug is administered to the patient, whereby the genetically engineered T-cells are killed, thereby preventing graft-versus-host disease.
  • a method of preventing graft-versus-host disease in a host that is being treated for a disease or disorder which is treatable by administering T-cells to a host such as, for example, a relapsed or persistent leukemia.
  • the method comprises administering to a host T- cells genetically engineered to include a polynucleotide encoding a negative selective marker or "suicide" gene.
  • the cells are administered in an amount effective and remain in the host for a period of time effective to provide a therapeutic effect in the host.
  • an interaction agent or prodrug is administered to the host.
  • the interaction agent is administered to the host in an amount effective to kill the genetically engineered T-cells, in particular, those T-cells which are graft-versus-host reactive, i.e., capable of providing a graft-versus-host effect, thereby preventing the occurrence of graft-versus-host disease in the host.
  • polynucleotide as used herein means a polymeric form of nucleotide of any length, and includes ribonucleotides and deoxyribonucleotides. Such term also includes single- and double-stranded DNA, as well as single- and double-stranded RNA. The term also includes modified polynucleotides such as methylated or capped polynucleotides.
  • the polynucleotide encoding the negative selective marker may be contained within an appropriate expression vehicle which is transduced into the T-cells.
  • expression vehicles include, but are not limited to, eukaryotic vectors, prokaryotic vectors (such as, for example, bacterial vectors), and viral vectors.
  • the polynucleotide encoding the agent or an expression vehicle containing the polynucleotide encoding the agent, is contained within a liposome.
  • the expression vehicle is a viral vector.
  • Viral vectors which may be employed include DNA virus vectors (such as adenoviral vectors, adeno- associated virus vectors, Herpes Virus vectors, and vaccinia virus vectors) , and RNA virus vectors (such as retroviral vectors) .
  • DNA virus vectors such as adenoviral vectors, adeno- associated virus vectors, Herpes Virus vectors, and vaccinia virus vectors
  • RNA virus vectors such as retroviral vectors
  • the viral vector is a retroviral vector.
  • retroviral vectors which may be employed include, but are not limited to, Moloney Murine Leukemia Virus, spleen necrosis virus, and vectors derived from retroviruses such as Rous Sarcoma Virus, Harvey Sarcoma Virus, avian leukosis virus, human immunodeficiency virus, myeloproliferative sarcoma virus, and mammary tumor virus.
  • the vector is preferably an infectious, replication incompetent retrovirus particle.
  • Retroviral vectors are useful as agents to mediate retroviral-mediated gene transfer into eukaryotic cells. Retroviral vectors are generally constructed such that the majority of sequences coding for the structural genes of the virus are deleted and replaced by the gene(s) of interest. Most often, the structural genes (i.e., gag, pol, and env) , are removed from the retroviral backbone using genetic engineering techniques known in the art.
  • a vector backbone comprised of a 5' LTR, a packaging signal, one or more cloning sites, into which the heterologous gene or genes of interest can be introduced, and a 3' LTR.
  • the preferred vector backbone is the Gl vector backbone, which is disclosed in McLachlin, et al., Virology. 195:1-5 (1993) and in PCT Patent Application No. WO 91/10728 for "Novel Retroviral Vectors,” published on July 25, 1991.
  • the heterologous gene or genes are incorporated into the proviral backbone by standard techniques to form the retroviral vector.
  • Techniques for the preparation of retroviral vectors are disclosed in PCT application WO 91/10728 as well as the following articles: Armentano, et al., J. Virol.. 61:1647-1650 (1987), Bender, et al. , J. Virol.. 61:1639-1646 (1987), and Miller, et al. , Biotechni ⁇ ues. 7:980-990 (1989) .
  • the most straightforward constructions are ones in which the structural genes of the retrovirus are replaced by a single gene which then is transcribed under the control of the viral regulatory sequences within the long terminal repeat (LTR) .
  • LTR long terminal repeat
  • Retroviral vectors have also been constructed which can introduce more than one gene into target cells.
  • one gene is under the regulatory control of the viral LTR, while the second gene is expressed either off a spliced message or is under the regulation of its own, internal promoter.
  • Suitable promoters include the SV40 promoter, the human cytomegalovirus (CMV) promoter, the beta-actin promoter, the alpha fetoprotein promoter, and any promoter naturally associated with any heterologous gene of interest.
  • CMV human cytomegalovirus
  • beta-actin promoter the beta-actin promoter
  • alpha fetoprotein promoter the alpha fetoprotein promoter
  • a polycistronic vector can be created by using an internal ribosome entry site.
  • the retroviral vectors may be in the form of a plasmid, a segment of viral RNA, or a segment of proviral DNA.
  • the preferred retroviral vector is GlTKlSvNa, which is disclosed in PCT Patent Application No. WO 95/06486, published on March 9, 1995, entitled Treatment of Human Tumors by Genetic Transformation of Human Tumor Cells.
  • the retroviral vector is introduced into a packaging cell to form a producer cell.
  • Packaging cells provide the gag, pol, and env genes in trans, which permits the packaging of the retroviral vector into a recombinant retrovirus that is infectious but replication defective.
  • the vectors are transferred into the packaging cells by standard gene transfer techniques, which include transfection, transduction, calcium phosphate precipitation, electroporation, and liposome-mediated DNA transfer.
  • packaging cells examples include, but are not limited to, the PE501, PA317, Psi-2, Psi-AM, PA12, T19- 14X, VT-19-17-H2, Psi-CRE, Psi-CRIP, GP+E-86, GP+envAM12, PG13, and DAN cell lines.
  • a preferred producer cell line for the present invention for the production of recombinant retrovirus is the producer cell line designated PA317/GlTKlSvNa, which is disclosed in PCT application WO 95/06486.
  • Negative selective markers include, but are not limited to, viral thymidine kinases such as Herpes Simplex Virus thymidine kinase, cytomegalovirus thymidine kinase, and varicella-zoster virus thymidine kinase; xanthine-guanine phosphoribosyl transferase; and cytosine deaminase.
  • viral thymidine kinases such as Herpes Simplex Virus thymidine kinase, cytomegalovirus thymidine kinase, and varicella-zoster virus thymidine kinase
  • xanthine-guanine phosphoribosyl transferase such as cytosine deaminase.
  • the retroviral vectors containing the polynucleotide encoding the negative selective marker are transduced into T- cells.
  • T-cells are transduced with the retroviral vectors, which may be contained in from about 2 ml to about 500 ml of retroviral supernatant having a titer of from about 10 5 cfu/ml to about 10 9 cfu/ml, preferably from about 2xl0 6 cfu/ml to about lxlO 8 cfu/ml.
  • the T-cells are administered to a host suffering from a relapsed or chronic leukemia.
  • the host is an animal host, and in particular is a mammalian host, including human and non-human primate hosts.
  • the transduced T-cells are administered by means known to those skilled in the art, including intravascular administration, such as intravenous or intraarterial administration; or by intraperitoneal administration.
  • the transduced T-cells are administered as a bolus infusion.
  • the transduced T-cells are administered in an amount effective to provide a therapeutic effect, i.e., in an amount effective to treat the relapsed or persistent leukemia in the host.
  • the transduced T-cells are administered in an amount of from about 10 5 cells/kg to about 10 9 cells/kg, preferably from about 2xl0 f cells/kg to about lxlO 7 cells/kg.
  • the transduced T-cells are administered in conjunction with an acceptable pharmaceutical carrier, such as, for example, saline solution, or aqueous buffers, such as phosphate buffers, Tris buffers, Plasmalyte A (Baxter) , or lactated Ringer's solution.
  • an acceptable pharmaceutical carrier such as, for example, saline solution, or aqueous buffers, such as phosphate buffers, Tris buffers, Plasmalyte A (Baxter) , or lactated Ringer's solution.
  • a suitable pharmaceutical carrier is deemed to be apparent to those skilled in the art from the teachings contained herein.
  • the transduced T-cells may be frozen in an acceptable cryopreservation medium (such as, for example, a medium including phosphate buffered saline, 5% DMSO, and human albumin) until the cells are administered to the host. Prior to administration, the cells and medium are thawed, and the cells and medium are administered to the host upon
  • an interaction or chemotherapeutic agent is administered to the host in an amount effective to kill the transduced T-cells, and in particular, to kill transduced proliferating graft-versus-host-reactive T-cells, i.e., T- cells which are capable of providing a graft-versus-host effect, thereby preventing the development of graft-versus- host disease.
  • the interaction agent is administered when the T-cells which are graft-versus-host reactive (whereby such T-cells which are capable of providing a graft-versus- host effect through the recognition of MHC Class I antigens of the host cells) are in a proliferative phase, and a portion of T-cells such as, for example, some of those T- cells which provide a graft-versus-leukemia effect or which provide an anti-viral effect, are in a quiescent or non- proliferative phase.
  • T-cells which are graft-versus-host reactive whereby such T-cells which are capable of providing a graft-versus- host effect through the recognition of MHC Class I antigens of the host cells
  • a portion of T-cells such as, for example, some of those T- cells which provide a graft-versus-leukemia effect or which provide an anti-viral effect
  • the interaction agent, or prodrug, when administered at such a time, will provide for the killing of the proliferating T-cells which are capable of providing a graft-versus-host effect, whereas the non-proliferating T- cells, which include a portion of T-cells which provide a graft-versus-leukemia effect or an anti-viral effect, will survive the administration of the interaction agent or prodrug.
  • the majority of the T-cells responsible for causing GVHD are ablated preferentially, while other T-cells capable of providing a graft-versus-leukemia effect or an anti-viral effect remain.
  • the interaction agent is administered at a period of time of from about 10 days to about 50 days, preferably from about 14 days to about 28 days, more preferably at 21 days, after the administration of the transduced T-cells.
  • the interaction or chemotherapeutic agent or prodrug preferably is a nucleoside analogue, for example, one selected from the group consisting of ganciclovir, acyclovir, and l-2-deoxy-2-fluoro- jS-D-arabinofuranosil-5-iodouracil (FIAU) .
  • FIAU l-2-deoxy-2-fluoro- jS-D-arabinofuranosil-5-iodouracil
  • Such interaction agents are utilized efficiently by the viral thymidine kinases as substrates, and such interaction agents thus are incorporated lethally into the DNA of the transduced T-cells expressing the viral thymidine kinases, and in particular, proliferating T-cells which are graft-versus-host reactive, thereby resulting in the death of the transduced T-cells.
  • cytosine deaminase When the negative selective marker is cytosine deaminase, a preferred interaction agent or prodrug is 5- fluorocytosine. Cytosine deaminase converts 5-fluorocytosine to 5-fluorouracil, which is highly cytotoxic. Thus, the transduced T-cells which express the cytosine deaminase gene convert the 5-fluorocytosine to 5-fluorouracil and are killed.
  • the interaction agent or prodrug is administered in an amount effective to provide for the death of the transduced T-cells.
  • the interaction agent is administered preferably by systemic administration, such as by intravenous administration.
  • the interaction agent is administered in an amount of from about 2 mg/kg/day to about 10 mg/kg/day, preferably about 10 mg/kg/day, for a period of from about 3 to 18 days, preferably for about 5 days.
  • the interaction agent or prodrug is administered in an amount of 5 mg/kg every 12 hours for a period of 5 days.
  • the interaction agent is administered upon the development of the graft-versus-host disease, thereby treating the graft-versus-host disease in the host.
  • the method of the present invention enables one to treat a patient suffering from a relapsing or chronic hematological malignancy or leukemia by administering T-cells to the patient, and to prevent the occurrence of graft- versus-host disease by killing the T-cells after the T-cells have provided a desired therapeutic effect.
  • Leukemias which may be treated with the transduced T-cells include, but are not limited to, multiple myeloma (MM) , myelodysplastic syndrome, chronic myelogenous leukemia (CML) , chronic lymphocytic leukemia (CLL) , acute myeloid leukemia (AMD , acute lymphoblastic leukemia (ALL), non-Hodgkin's lymphoma, Hodgkin's disease, and myelofibrosis.
  • MM multiple myeloma
  • CML chronic myelogenous leukemia
  • CLL chronic lymphocytic leukemia
  • AMD acute myeloid leukemia
  • ALL acute lymphoblastic leukemia
  • non-Hodgkin's lymphoma Hodgkin's disease
  • myelofibrosis myelofibrosis
  • An advantage of the present invention is that the T- cells which are genetically engineered to include a polynucleotide encoding a negative selective marker are safer than T-cells which are not genetically engineered to be employed in the treatment of relapsed or chronic leukemia because the major cause of mortality and morbidity, graft- versus-host disease, may be prevented by the administration of a prodrug or interaction agent. Because graft-versus-host disease is a consequence of the degree of mismatch between the donor and patient, the method of the present invention expands the number of patients that are eligible for an allogeneic bone marrow transplant to include older patients and more genetically disparate donors, including HLA non- identical siblings and matched, unrelated donors.
  • the timing of the administration of the interaction agent or prodrug is such that the majority of the GVHD reactive cells (i.e., the T-cells that are responsible for causing GVHD) are ablated preferentially, thus leaving the patient with the greater part of the immunologic T-cell repertoire. It is likely that at least a part of the graft- versus-leukemia reactive cells and T-cells which are reactive with antigens other than those associated with GVHD, will survive the administration of the interaction agent or prodrug, thus helping to decrease the frequency of EBV lymphomas and infections.
  • the method hereinabove described may be applied to the treatment of other diseases and disorders.
  • the method may be applied to the treatment of solid tumor malignancies, especially in cases where the tumor has metastasized and tumor cells are found in the bone marrow.
  • patients would undergo a high dose chemotherapy treatment with or without total body irradiation followed by rescue with an allogeneic T-cell depleted bone marrow transplant.
  • T-cells genetically engineered with a polynucleotide encoding a negative selective marker or suicide gene would be administered to the patient as hereinabove described, and GVHD would be prevented by the administration of the interaction agent or prodrug.
  • the T-cells provide a graft-versus-tumor (GVT) effect.
  • Solid tumor malignancies which may be treated in accordance with this method include, but are not limited to, breast cancer, neuroblastoma, testicular carcinoma, ovarian and uterine carcinomas, and soft tissue sarcomas.
  • the hereinabove described method of the present invention may be employed in the treatment of any acquired or genetic immunologic or hematopoietic disease in which treatment thereof includes bone marrow ablation followed by the administration of a T-cell depleted bone marrow transplant.
  • Such diseases include, but are not limited to, AIDS, severe combined immune deficiency (SCID) , Wiskott-Aldrich syndrome, disorders of lymphocyte function, disorders of myeloid function, aplastic anemia, Fanconi' ⁇ anemia, thalassemia, sickle cell anemia, enzyme deficiencies (including Gaucher's disease, the mucopolysaccharidoses, and the leukodystrophies) , and osteopetrosis, a disease which is characterized by a deficiency in bone marrow cells which break down older bone.
  • the genetically engineered T-cells in this embodiment, would be responsible for improved bone marrow engraftment, and for decreasing the incidence of EBV lymphoma.
  • GVHD is prevented by administration of the prodrug or interaction agent.
  • a method of treating a disease or disorder in a host wherein treatment of the disease or disorder in the host includes ablating the bone marrow of the host, followed by the administration of a T-cell depleted bone marrow transplant to the host.
  • the method comprises ablating the bone marrow of the host.
  • a T-cell depleted bone marrow transplant then is administered to the host.
  • the host then is administered T- cells genetically engineered to include a polynucleotide encoding a negative selective marker, which may be selected from those hereinabove described.
  • an interaction agent or prodrug is administered to the host in an amount effective to kill the genetically engineered T-cells in the host.
  • polynucleotide encoding the negative selective marker may be contained in an appropriate expression vehicle such as those hereinabove described, including viral vectors such as retroviral vectors.
  • the genetically engineered T-cells may be administered to the host in an amount effective to provide a therapeutic effect. Such amount may be as hereinabove described.
  • the exact amount of genetically engineered T-cells to be administered is dependent upon a variety of factors, including the age, weight, and sex of the patient, the disease or disorder being treated, and the extent and severity thereof.
  • the prodrug or interaction agent is administered in an amount, which may be as hereinabove described, which is effective in killing the genetically engineered T-cells, and in particular, those genetically engineered T-cells capable of providing a graft-versus-host effect, thereby preventing the development of graft-versus- host disease.
  • the period of time after the administration of the genetically engineered T-cells at which the interaction agent is administered may be as hereinabove described.
  • T-cells transduced with a retroviral vector including a Herpes Simplex Virus thymidine kinase gene to patients suffering from persistent or relapsing multiple myeloma, followed by administration of ganciclovir
  • ⁇ blood 50 to 100 ml of donor blood are collected, yielding at least 2.5-5xl0 7 cells.
  • the blood is mixed with an equal volume of sterile phosphate buffered saline (PBS) and subjected to gradient separation using Ficoll (Lymphoprep, Nycomed) , and the mononuclear cells are separated. The cells then are washed in sterile PBS three times.
  • PBS sterile phosphate buffered saline
  • the cells are resuspended in serum-free AIM-V medium containing 100 units/ml penicillin and 100 ⁇ g/ml streptomycin (Pen-Strep, Gibco-BRL) at a density of lxlO 6 cells/ml with l ⁇ g/ml anti-CD3 antibody (Orthoclone OKT3, Ortho-Biotech), and incubated at 37°C in 5% C0 2 for 24 hours.
  • Recombinant human Interleukin-2 (Chiron Corporation) is added at 1,500 units/ml and cells are cultured at 37°C in 5% C0 2 for a period of up to 7 days.
  • the lymphocytes are in the exponential proliferation phase, they are transduced with the retroviral vector.
  • Approximately 1 X 10 8 lymphocytes are transduced with the retroviral vector GlTKlSvNa, which includes the Herpes Simplex Virus thymidine kinase (TK) gene and a neomycin resistance gene.
  • TK Herpes Simplex Virus thymidine kinase
  • neomycin resistance gene a gene that confers the ability to transduction.
  • Such transduction is accomplished by adding to the lymphocytes 100 ml of viral supernatant containing from 2xl0 8 cfu to lxlO 9 cfu of the retroviral vector.
  • the retroviral vector GlTKlSvNa is described further in PCT Application No. WO95/06486, published March 9, 1995.
  • Fresh viral supernatant is added daily for a total of three days, along with additional amounts of Interleukin - 2 and protamine sulfate to achieve final Interleukin - 2 concentrations of 1,500 units/ml and protamine sulfate at 5 ⁇ g/ml.
  • the cells are incubated at 37°C, 5% C0 2 for 24 hours after each addition of supernatant and protamine sulfate.
  • the cells are resuspended at a density of 1 X 10 6 cells/ml in fresh medium containing Pen-Strep and 1,500 units/ml of Interleukin-2 and incubated for 3 to 5 days. Geneticin then is added at an active concentration of 300 ⁇ g/ml, and cells are selected for three days, are centrifuged and then are resuspended in AIM-V medium containing Interleukin-2 and cultured until an adequate number of cells are obtained. In general, the cells are cultured for at least an additional four days. The efficiency of transduction and selection is measured by a ganciclovir killing assay. One to two percent of the transduced cells are sampled for the ganciclovir killing assay.
  • the assay is performed by suspending transduced lymphocytes in 5.0 ml of AIM-V (Gibco-BRL) medium containing 1,500 units/ml recombinant Interleukin-2 in tissue culture flasks. A total of three flasks are prepared. Ganciclovir is added to two of the flasks, one at a concentration of 20 ⁇ M and the second at 50 ⁇ M. The third flask is used as a normal control. The cells are incubated at 37°C, in 5% C0 2 for five days. The percentage of live cells (trypan blue negative) are counted after 3 and 5 days of culture in ganciclovir.
  • AIM-V Gibco-BRL
  • the cells are reselected in Geneticin as described above. Cells are cryopreserved and infused only if the ganciclovir kill is greater than or equal to 85%. In order to provide enough cells for two infusions (if needed) , an adequate number of cells initially is collected, expanded, and cryopreserved for each patient.
  • the transduced lymphocytes also are studied for subset analysis (CD3, CD4, CD8, CD19, and CD56) by FACS prior to infusion. Infusion of transduced lymphocytes and administration of ganciclovir.
  • Infusion of the transduced lymphocytes is given to patients who have undergone an allogeneic bone marrow transplant with T-cell depletion if they show evidence of persistent disease 90 days after transplantation, or measurable relapse at anytime.
  • Patients with persistent disease have greater than 20% plasma cells in a bone marrow aspirate or biopsy, and/or presence of serum M-component, and no reduction in the M-component in the last 6 weeks, and/or Bence Jones Proteinuria with no reduction in the last 6 weeks.
  • Each of three patients receives the transduced lymphocytes in an amount of 1 X 10 6 lymphocytes/kg.
  • One to 50 ml of lymphocytes is administered as a bolus infusion, and each patient is observed for 4 hours thereafter.
  • GVHD graft-versus-host disease
  • Complete response should include, for a minimum of two weeks, the following: (i) absence in urine and serum of M- components by immunofixation; (ii) bone marrow which is adequately cellular (i.e. >20%) with less than 3% plasma cells by immunostaining; (iii) no elevation in serum calcium level; and (iv) no new bone lesions nor enlargement of existing lesions.
  • Partial response requires, for at least 4 weeks, the following: (i) reduction of serum M-component by at least 5%; (ii) reduction of urinary M-protein to less than 200 mg/24 hrs. and to less than 10% of pretreatment values; and (iii) no new lytic bone lesions or soft tissue plasmacytoma. Improvement also should include a reduction of serum paraprotein level and urinary light chain excretion by 25-50% compared with baseline values, and a decrease of bone marrow infiltration by plasma cells by 25-50% compared with baseline values.
  • dose levels are as follows:
  • GVHD is graded as follows:
  • the dose of lymphocytes is escalated from 1 X 10 6 lymphocytes/kg to 5 X 10 6 lymphocytes/kg (i.e., dose level ID .
  • the treatment plan for each dose level is identical to the one described above for dose level I (i.e., l X 10 6 lymphocytes/kg) .
  • Six additional patients are entered at the dose level at which one of three patients responds, or at one dose below the level at which toxicity is observed.

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Abstract

Cette invention concerne un procédé de prévention de la réaction du greffon contre l'hôte chez un patient traité pour leucémie persistante ou récurrente. Ledit procédé consiste à administrer à un hôte des lymphocytes thymo-dépendants produits par génie génétique de façon à incorporer un polynucléotide codant un marqueur de sélection négative. Avant l'apparition de la réaction du greffon contre l'hôte, on administre à l'hôte un agent d'interaction ou un agent chimiothérapeutique qui détruit les lymphocytes thymo-dépendants produits par génie génétique et empêche l'apparition de ladite réaction. On peut mettre en oeuvre ce procédé parallèlement au traitement de toute maladie ou trouble impliquant une ablation de moelle osseuse suivie d'une greffe de moelle osseuse appauvrie en lymphocytes thymo-dépendants.
PCT/US1997/009040 1996-05-31 1997-05-28 Prevention de la reaction du greffon contre l'hote au moyen de lymphocytes thymo-dependants contenant des polynucleotides codant des marqueurs de selection negative WO1997045142A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
IL12721197A IL127211A0 (en) 1996-05-31 1997-05-28 Prevention of graft-versus-host disease with t-cells including polynucleotides encoding negative selective markers
AU30796/97A AU719930B2 (en) 1996-05-31 1997-05-28 Prevention of graft-versus-host disease with T-cells including polynucleotides encoding negative selective markers
JP09542889A JP2000511539A (ja) 1996-05-31 1997-05-28 負の選択マーカーをコード化するポリヌクレオチドを含むｔ細胞による対宿主性移植片病の予防
EP97925747A EP0977595A4 (fr) 1996-05-31 1997-05-28 Prevention de la reaction du greffon contre l'hote au moyen de lymphocytes thymo-dependants contenant des polynucleotides codant des marqueurs de selection negative
NZ333053A NZ333053A (en) 1996-05-31 1997-05-28 Use of an interaction agent or T-cells having polynucleotides encoding negative selective markers to prevent graft-versus-host disease
NO985522A NO985522L (no) 1996-05-31 1998-11-26 Forebyggelse av graft-versus-host sykdom med T-celler innbefattende polynukleotider som koder negative selektive mark÷rer

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US65645196A 1996-05-31 1996-05-31
US08/656,451 1996-05-31

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WO1997045142A1 true WO1997045142A1 (fr) 1997-12-04

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EP (1) EP0977595A4 (fr)
JP (1) JP2000511539A (fr)
AU (1) AU719930B2 (fr)
CA (1) CA2255941A1 (fr)
IL (1) IL127211A0 (fr)
NO (1) NO985522L (fr)
NZ (1) NZ333053A (fr)
WO (1) WO1997045142A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998042824A2 (fr) * 1997-03-20 1998-10-01 Cellfactors Plc Procedes permettant de selectionner des cellules et utilisations de ces procedes
FR2780891A1 (fr) * 1998-07-10 2000-01-14 Univ Paris Curie Echange de lymphocytes t
WO2000076542A1 (fr) * 1999-06-11 2000-12-21 Consiglio Nazionale Delle Ricerche Utilisation d'anticorps contre cd20 pour traiter la maladie de rejet du greffon
WO2007054183A1 (fr) * 2005-11-10 2007-05-18 Schering Aktiengesellschaft Réduction de resténose
US7452723B2 (en) 1997-08-13 2008-11-18 Oncolytics Biotech Inc. Methods for preventing reovirus recognition for the treatment of cellular proliferative disorders
US7582289B2 (en) 1999-11-12 2009-09-01 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
EP2210608A1 (fr) * 2007-11-02 2010-07-28 JCR Pharmaceuticals CO., LTD. Composition pharmaceutique contenant des cellules-souches mésenchymateuses humaines
US9993550B2 (en) 1999-05-07 2018-06-12 Genentech, Inc. Treatment of pemphigus

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2456924C1 (ru) * 2010-12-14 2012-07-27 Федеральное государственное учреждение "Кировский научно-исследовательский институт гематологии и переливания крови Федерального медико-биологического агентства" Способ прогнозирования общей выживаемости больных множественной миеломой

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
BLOOD, 01 August 1996, Vol. 88, No. 3, GALLOT G. et al., "Human HLA-Specific T-Cell Clones with Stable Expression of a Suicide Gene: A Possible Tool to Drive and Control a Graft-Versus-Host--Graft-Versus-Leukemia Reaction?", pages 1098-1103. *
BLOOD, 1994, Abstract 427, BONINI C. et al., "Transfer of the HSV-TK Gene Into Donor Peripheral Blood Lymphocytes for in Vivo Immunomodulation of Donor Anti-Tumor Immunity After Allo-BMT". *
BLOOD, November 1995, Abstract 843, SERVIDA P. et al., "Gene Transfer Into Peripheral Blood Lymphocytes for in Vivo Immunomodulation of Donor Anti-Tumor Immunity in a Patient Affected by EBV-Induced Lymphoma". *
NIH OFFICE OF PUBLIC INFORMATION, 7 December 1995, ORKIN S.H. et al., "Report and Recommendations of the Panel to Assess the NIH Investment in Research on Gene Therapy", pages 1-40. *
See also references of EP0977595A4 *

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998042824A3 (fr) * 1997-03-20 1999-01-07 Cellfactors Plc Procedes permettant de selectionner des cellules et utilisations de ces procedes
WO1998042824A2 (fr) * 1997-03-20 1998-10-01 Cellfactors Plc Procedes permettant de selectionner des cellules et utilisations de ces procedes
US7708987B2 (en) 1997-08-13 2010-05-04 Oncolytics Biotech Inc. Methods for preventing reovirus recognition for the treatment of cellular proliferative disorders
US7452723B2 (en) 1997-08-13 2008-11-18 Oncolytics Biotech Inc. Methods for preventing reovirus recognition for the treatment of cellular proliferative disorders
FR2780891A1 (fr) * 1998-07-10 2000-01-14 Univ Paris Curie Echange de lymphocytes t
EP0972518A2 (fr) * 1998-07-10 2000-01-19 Universite Pierre Et Marie Curie Paris Vi Echange de lymphocytes T
WO2000002520A2 (fr) * 1998-07-10 2000-01-20 Universite Pierre Et Marie Curie (Paris Vi) Echange de lymphocytes t
EP0972518A3 (fr) * 1998-07-10 2000-01-26 Universite Pierre Et Marie Curie Paris Vi Echange de lymphocytes T
WO2000002520A3 (fr) * 1998-07-10 2000-03-23 Univ Paris Curie Echange de lymphocytes t
US9993550B2 (en) 1999-05-07 2018-06-12 Genentech, Inc. Treatment of pemphigus
WO2000076542A1 (fr) * 1999-06-11 2000-12-21 Consiglio Nazionale Delle Ricerche Utilisation d'anticorps contre cd20 pour traiter la maladie de rejet du greffon
US7582289B2 (en) 1999-11-12 2009-09-01 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US7731951B2 (en) 1999-11-12 2010-06-08 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US7799329B2 (en) 1999-11-12 2010-09-21 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US7964186B2 (en) 1999-11-12 2011-06-21 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US8080241B2 (en) 1999-11-12 2011-12-20 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US8491886B2 (en) 1999-11-12 2013-07-23 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
WO2007054183A1 (fr) * 2005-11-10 2007-05-18 Schering Aktiengesellschaft Réduction de resténose
EP2210608A1 (fr) * 2007-11-02 2010-07-28 JCR Pharmaceuticals CO., LTD. Composition pharmaceutique contenant des cellules-souches mésenchymateuses humaines
EP2210608A4 (fr) * 2007-11-02 2013-03-20 Japan Chem Res Composition pharmaceutique contenant des cellules-souches mésenchymateuses humaines
JP5394932B2 (ja) * 2007-11-02 2014-01-22 日本ケミカルリサーチ株式会社 ヒト間葉系幹細胞含有医薬組成物

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IL127211A0 (en) 1999-09-22
JP2000511539A (ja) 2000-09-05
EP0977595A1 (fr) 2000-02-09
AU719930B2 (en) 2000-05-18
NO985522L (no) 1999-01-25
EP0977595A4 (fr) 2001-05-16
AU3079697A (en) 1998-01-05
NZ333053A (en) 2000-06-23
CA2255941A1 (fr) 1997-12-04
NO985522D0 (no) 1998-11-26

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