Claims
- 1. A method of distinguishing between heparanase expressing hematopoietic cells and heparanase non-expressing hematopoietic cells in subject in need thereof, the method comprising monitoring heparanase expression in hematopoietic cells obtained from the subject and, based on a presence or an absence of said heparanase expression, distinguishing between said heparanase expressing hematopoietic cells and said heparanase non-expressing hematopoietic cells.
- 2. The method of claim 1, wherein said hematopoietic cells are peripheral blood cells.
- 3. The method of claim 1, wherein said hematopoietic cells are bone marrow cells.
- 4. The method of claim 1, wherein said hematopoietic cells are lymph node cells.
- 5. The method of claim 1, wherein said heparanase expressing hematopoietic cells comprise activated 1 or B lymphocytes, platelets, granulocytes, macrophages, monocytes, mast cells, megakaryocytes, promegakaryocytes, megakaryoblasts, promyelocytes, metamyelocytes, myelocytes, myeloblasts, monoblasts and dendritic cells.
- 6. The method of claim 1, wherein said heparanase non-expressing hematopoietic cells comprise hematopoietic stern cells, erythrocytes, normoblasts, erythroblasts naive T and B lymphocytes.
- 7. The method of claim 1, wherein said hematopoietic cells additionally express the surface markers CD2, CD3, CD4, CD7, CD8, CD10, CD11b, CD13 CD14, CD15, CD16, CD18, CD19, CD20 CD22, CD23, CD25, CD28, CD33, CD34, CD38, CD41, CD42a, CD42b, CD45, CD61, CD71, CD103, CD117, HLA-DR, MPO, FMC7 and TdT.
- 8. The method of claim 1, used to distinguish between heparanase expressing myeloid cells and heparanase non-expressing B lymphoma cells.
- 9. The method of claim 1, used to distinguish between heparanase expressing myeloid cells (in acute myeloid leukemia) and heparanase non-expressing myeloid cells (in chronic myeloid leukemia)
- 10. The method of claim 1, used to distinguish between heparanase expressing myeloid cells and heparanase non-expressing B leukemia cells.
- 11. The method of claim 1, used to distinguish between heparanase expressing acute myelogenous leukemia cells and heparanase non-expressing chronic lymphocytic leukemia cells.
- 12. The method of claim 1, used to distinguish between heparanase expressing acute myelogenous leukemia cells and heparanase non-expressing non-Hodgkin's lymphoma cells.
- 13. The method of claim 1, used to distinguish between heparanase expressing acute myelogenous leukemia cells and heparanase non-expressing chronic myeloid leukemia cells.
- 14. The method of claim 1, used to distinguish between heparanase expressing acute myelogenous leukemia cells and heparanase non-expressing Acute lymphocytic leukemia cells.
- 15. The method of claim 1, wherein monitoring said heparanase expression is by monitoring heparanase RNA levels.
- 16. The method of claim 15, wherein monitoring said heparanase RNA level is effected by an assay selected from the group consisting of RT-PCR, chip hybridization, RNase protection, in-situ hybridization, primer extension, Northern blot and dot blot analysis.
- 17. The method of claim 1, wherein monitoring said heparanase expression is by monitoring heparanase protein level.
- 18. The method of claim 17, wherein monitoring said heparanase protein level is effected by an assay selected from the group consisting of immunohistochemistry, ELISA, RIA, Western blot analysis, FACS analysis, an immunofluorescence assay, and a light emission immunoassay.
- 19. The method of claim 1, wherein monitoring said heparanase expression is by monitoring heparanase activity level.
- 20. The method of claim 9, wherein said monitoring said heparanase activity level is effected by using a heparanase substrate.
- 21. The method of claim 20, wherein said heparanase substrate is selected from the group consisting of soluble or immobilized heparan sulfate proteoglycans, heparan sulfate, or heparin.
- 22. The method of claim 20, wherein said heparanase substrate includes a detectable moiety selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a radioactive moiety and a light-emitting moiety.
- 23. The method of claim 1, wherein said hematopoietic cells obtained from said subject are further enriched for leukocytes prior to said monitoring said heparanase expression.
- 24. A kit for distinguishing between heparanase expressing hematopoietic cells and heparanase non-expressing hematopoietic cells, comprising at least one container including at least one reagent for determining a level of heparanase expression or activity in a biological sample, and packaging material identifying said at least one reagent for use in distinguishing between heparanase expressing hematopoietic cells and heparanase non-expressing hematopoietic cells.
- 25. The kit of claim 24, wherein said heparanase hematopoietic cells and/or said heparanase non-expressing hematopoietic cells are isolated from a bone marrow, a lymph node or peripheral blood.
- 26. The kit of claim 24, wherein distinguishing between heparanase expressing hematopoietic cells and heparanase non-expressing hematopoietic cells is by monitoring heparanase RNA levels within said cells, said at least one reagent comprises a substance selected for specifically interacting with heparanase mRNA.
- 27. The kit of claim 26, wherein said monitoring said heparanase RNA levels is effected by an assay selected from the group consisting of RT-PCR, chip hybridization, RNase protection, in-situ hybridization, primer extension, Northern blot and dot blot analysis, said at least one reagent comprises a substance for effecting said assay.
- 28. The kit of claim 26, wherein said at least one reagent for determining a level of heparanase expression in a biological sample comprises oligonucleotide primers for the identification and/or amplification of heparanase mRNA.
- 29. The kit of claim 24, wherein distinguishing between heparanase expressing hematopoietic cells and heparanase non-expressing hematopoietic cells is by monitoring heparanase protein level or activity in said cells, said at least one reagent is selected for interacting with heparanase.
- 30. The kit of claim 29, wherein said monitoring said heparanase protein level is effected by an assay selected from the group consisting of immunohistochemistry, ELISA, RIA, Western blot analysis, FACS analysis, an immunofluorescence assay, and a light emission immunoassay, said at least one reagent comprises a heparanase specific antibody.
- 31. The kit of claim 29, wherein said at least one reagent comprises a heparanase specific antibody.
- 32. The kit of claim 31, wherein said heparanase specific antibody is coupled to an enzyme.
- 33. The kit of claim 29, wherein said at least one reagent comprises a heparanase substrate.
- 34. The kit of claim 33, wherein said heparanase substrate is selected from the group consisting of soluble or immobilized heparan sulfate proteoglycans, heparan sulfate or heparin.
- 35. The kit of claim 33, wherein said heparanase substrate comprises a detectable moiety selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a radioactive moiety and a light-emitting moiety.
- 36. An improvement in a process of diagnosing acute myelogenous leukemia, the improvement comprising determining whether cells suspected as leukemic cells express heparanase, wherein heparanase expression is indicative of acute myelogenous leukemia.
- 37. The improvement of claim 36, wherein said suspected leukemic cells are obtained from a bone marrow, a lymph node or peripheral blood.
- 38. The improvement of claim 36, wherein determining heparanase expression is by monitoring heparanase RNA level.
- 39. The improvement of claim 38, wherein monitoring said heparanase RNA level is effected by an assay selected from the group consisting of RT-PCR, chip hybridization, RNase protection, in-situ hybridization, primer extension, Northern blot and dot blot analysis.
- 40. The improvement of claim 36, wherein determining heparanase expression is by monitoring heparanase protein level.
- 41. The improvement of claim 40, wherein monitoring said heparanase protein level is effected by an assay selected from the group consisting of immunohistochemistry, ELISA, RIA, Western blot analysis, FACS analysis, an immunofluorescence assay, and a light emission immunoassay.
- 42. The improvement of claim 36, wherein determining heparanase expression is by monitoring heparanase activity level.
- 43. The improvement of claim 42, wherein said monitoring said heparanase activity level is effected by using a heparanase substrate.
- 44. The improvement of claim 43, wherein said heparanase substrate is selected from the group consisting of soluble or immobilized heparan sulfate proteoglycans, heparan sulfate or heparin.
- 45. The improvement of claim 43, wherein said heparanase substrate includes a detectable moiety selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a radioactive moiety and a light-emitting moiety.
- 46. A method for ascertaining the suitability of an anti-neoplastic drug candidate for efficacy in treating acute myelogenous leukemia, wherein said method comprises combining said candidate drug with a cell having an acute myelogenous leukemia phenotype, said cell being isolated or in a biological sample and detecting in said cell or biological sample a change in heparanase expression.
- 47. The method of claim 46, wherein said cell having an acute myelogenous leukemia phenotype is obtained from a bone marrow, a lymph node or peripheral blood.
- 48. The method of claim 46, wherein detecting said change in heparanase expression in said cell or biological sample is by monitoring heparanase RNA levels.
- 49. The method of claim 48, wherein monitoring said heparanase RNA levels is effected by an assay selected from the group consisting of RT-PCR, chip hybridization, RNase protection, in-situ hybridization, primer extension, Northern blot and dot blot analysis.
- 50. The method of claim 46, wherein detecting said change in heparanase expression in said cell or biological sample is by monitoring heparanase protein levels.
- 51. The method of claim 50, wherein monitoring said heparanase protein level is effected by an assay selected from the group consisting of immunohistochemistry, ELISA, RIA, Western blot analysis, FACS analysis, an immunofluorescence assay, and a light emission immunoassay.
- 52. The method of claim 46, wherein detecting said change in heparanase expression in said cell or biological sample is by monitoring heparanase activity level.
- 53. The method of claim 52, wherein said monitoring said heparanase activity level is effected by using a heparanase substrate.
- 54. The method of claim 53, wherein said heparanase substrate is selected from the group consisting of soluble or immobilized heparan sulfate proteoglycans, heparan sulfate or heparin, 57. The method of claim 53, wherein said heparanase substrate includes a detectable moiety selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a radioactive moiety and a light-emitting moiety.
- 55. A kit for distinguishing between heparanase expressing acute myeloid leukemic cells and heparanase non-expressing neoplastic or normal hematopoietic cells, comprising at least one container including at least one reagent for determining a level of heparanase expression or activity in a biological sample, and packaging material identifying said at least one reagent for use in distinguishing between heparanase expressing acute myeloid leukemic cells and heparanase non-expressing neoplastic or normal hematopoietic cells.
- 56. The kit of claim 55, wherein said heparanase expressing acute myeloid leukemic cells and/or said heparanase non-expressing neoplastic or normal hematopoietic cells are isolated from a bone marrow, a lymph node or peripheral blood.
- 57. The kit of claim 55, wherein distinguishing between heparanase expressing acute myeloid leukemic cells and heparanase non-expressing neoplastic or normal hematopoietic cells is by monitoring heparanase RNA levels within said cells, said at least one reagent comprises a substance selected for specifically interacting with heparanase mRNA.
- 58. The kit of claim 57, wherein monitoring said heparanase RNA levels is effected by an assay selected from the group consisting of RT-PCR, chip hybridization, RNase protection, in-situ hybridization, primer extension, Northern blot and dot blot analysis.
- 59. The kit of claim 57, wherein said at least one reagent for determining a level of heparanase expression in a biological sample comprises oligonucleotide primers for the identification and/or amplification of the heparanase gene.
- 60. The kit of claim 55, wherein distinguishing between heparanase expressing acute myeloid leukemic cells and heparanase non-expressing neoplastic or normal hematopoietic cells is by monitoring heparanase protein levels or activity within said cells, said at least one reagent is selected for interacting with heparanase.
- 61. The kit of claim 60, wherein monitoring said heparanase protein level is effected by an assay selected from the group consisting of immunohistochemistry, ELISA, RIA, Western blot analysis, FACS analysis, an immunofluorescence assay, and a light emission immunoassay, said at least one reagent comprises a heparanase specific antibody.
- 62. The kit of claim 60, wherein said reagent includes a heparanase specific antibody.
- 63. The kit of claim 62, wherein said heparanase specific antibody is coupled to an enzyme.
- 64. The kit of claim 60, wherein said at least one reagent comprises a heparanase substrate.
- 65. The kit of claim 64, wherein said heparanase substrate is selected from the group consisting of soluble or immobilized heparan sulfate proteoglycans, heparan sulfate or heparin,
- 66. The kit of claim 64, wherein said heparanase substrate comprises a detectable moiety selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a radioactive moiety and a light-emitting moiety.
- 67. An improvement in a process of diagnosing relapse in acute lymphocytic leukemia, the improvement comprising determining whether cells suspected as leukemic cells express heparanase, wherein heparanase expression is indicative of relapse of acute lymphocytic leukemia.
- 68. The improvement of claim 67, wherein said suspected leukemic cells are obtained from a bone marrow, a lymph node or peripheral blood.
- 69. The improvement of claim 67, wherein determining heparanase expression is by monitoring heparanase RNA levels.
- 70. The improvement of claim 69, wherein monitoring said heparanase RNA levels is by a methodology selected from the group consisting of RT-PCR, chip hybridization, RNase protection, in-situ hybridization, primer extension, Northern blot and dot blot.
- 71. The improvement of claim 67, wherein determining heparanase expression is by monitoring heparanase protein level.
- 72. The improvement of claim 71, wherein monitoring said heparanase protein level is effected by an assay selected from the group consisting of immunohistochemistry, ELISA, RIA, western blot analysis, FACS analysis, an immunofluorescence assay, and a light emission immunoassay.
- 73. The improvement of claim 67, wherein determining heparanase expression is by monitoring heparanase activity level.
- 74. The improvement of claim 73, wherein said monitoring said heparanase activity level is effected by using a heparanase substrate.
- 75. The improvement of claim 74, wherein said heparanase substrate is selected from the group consisting of heparan sulfate proteoglycans, heparan sulfate, heparin, and heparin-sepharose.
- 76. The improvement of claim 74, wherein said heparanase substrate includes a detectable moiety selected from the group consisting of a chromogenic, a fluorogenic, a radioactive and a light-emitting moiety.
- 77. An improvement in a process of diagnosing chronic myelogenous leukemia, the improvement comprising determining whether cells suspected as leukemic cells express heparanase, wherein non-expression of heparanase is indicative of diagnosing chronic myelogenous leukemia.
- 78. The improvement of claim 77, wherein said suspected leukemic cells are obtained from a bone marrow, a lymph node or peripheral blood.
- 79. The improvement of claim 77, wherein determining heparanase expression is by monitoring heparanase RNA levels.
- 80. The improvement of claim 79, wherein monitoring said heparanase RNA levels is by a methodology selected from the group consisting of RT-PCR, chip hybridization, RNase protection, in-situ hybridization, primer extension, Northern blot and dot blot.
- 81. The improvement of claim 77, wherein determining heparanase expression is by monitoring heparanase protein level.
- 82. The improvement of claim 81, wherein monitoring said heparanase protein level is effected by an assay selected from the group consisting of immunohistochemistry, ELISA, RIA, western blot analysis, FACS analysis, an immunofluorescence assay, and a light emission immunoassay.
- 83. The improvement of claim 77, wherein determining heparanase expression is by monitoring heparanase activity level.
- 84. The improvement of claim 83, wherein said monitoring said heparanase activity level is effected by using a heparanase substrate.
- 85. The improvement of claim 84, wherein said heparanase substrate is selected from the group consisting of soluble or immobilized heparan sulfate proteoglycans, heparan sulfate, or heparin.
- 86. The improvement of claim 84, wherein said heparanase substrate includes a detectable moiety selected from the group consisting of a chromogenic, a fluorogenic, radioactive and a light-emitting moiety.
- 87. A kit for distinguishing between heparanase expressing neoplastic or normal hematopoietic cells and heparanase non-expressing chronic myeloid leukemic cells, comprising at least one container including at least one reagent for determining a level of heparanase expression or activity in a biological sample, and packaging material identifying said at least one reagent for use in distinguishing between heparanase expressing neoplastic or normal hematopoietic cells and heparanase non-expressing chronic myeloid leukemic cells.
- 88. The kit of claim 87, wherein said heparanase expressing neoplastic or normal hematopoietic cells and/or said heparanase non-expressing chronic myeloid leukemic cells are isolated from a bone marrow, a lymph node or peripheral blood.
- 89. The kit of claim 87, wherein distinguishing between heparanase expressing neoplastic or normal hematopoietic cells and heparanase non-expressing chronic myeloid leukemic cells is by monitoring heparanase RNA levels within said cells, said at least one reagent comprises a substance selected for specifically interacting with heparanase mRNA.
- 90. The kit of claim 89, wherein monitoring said heparanase RNA levels is effected by an assay selected from the group consisting of RT-PCR, chip hybridization, RNase protection, in-situ hybridization, primer extension, Northern blot and dot blot analysis.
- 91. The kit of claim 89, wherein said at least one reagent for determining a level of heparanase expression in a biological sample comprises oligonucleotide primers for the identification and/or amplification of the heparanase gene.
- 92. The kit of claim 87, wherein distinguishing between heparanase expressing neoplastic or normal hematopoietic cells and heparanase non-expressing chronic myeloid leukemic cells is by monitoring heparanase protein levels or activity within said cells, said at least one reagent is selected for interacting with heparanase.
- 93. The kit of claim 92, wherein monitoring said heparanase protein level is effected by an assay selected from the group consisting of immunohistochemistry, ELISA, RIA, Western blot analysis, FACS analysis, an immunofluorescence assay, and a light emission immunoassay, said at least one reagent comprises a heparanase specific antibody.
- 94. The kit of claim 92, wherein said reagent includes a heparanase specific antibody.
- 95. The kit of claim 94, wherein said heparanase specific antibody is coupled to an enzyme.
- 96. The kit of claim 92, wherein said at least one reagent comprises a heparanase substrate.
- 97. The kit of claim 96, wherein said heparanase substrate is selected from the group consisting of heparan sulfate proteoglycans, heparan sulfate, heparin, and heparin-sepharose.
- 98. The kit of claim 96, wherein said heparanase substrate comprises a detectable moiety selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a radioactive moiety and a light-emitting moiety.
Parent Case Info
[0001] This Application claims the benefit of priority from U.S. Provisional Patent Application No. 60/314,302, filed Aug. 24, 2001.
Provisional Applications (1)
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Number |
Date |
Country |
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60314302 |
Aug 2001 |
US |