Non-human transgenic mammals used as models for human pathologies originating from stem cells

Abstract

Transgenic non-human mammals that reproduce human pathologies of stem cell origins, such as chromosomal anomalies associated with chronic myeloid leukemia, B-cell acute lymphoblastic leukemia, T-cell acute or lymphoblastic leukemia, or with the migration of hematopoietic or embryonic stem calls. The transgenic non-human mammals can be produced using as a strategy the expression of genes involved in said pathologies by means of a promoter that directs the expression of a transgene in Sca-1 + cells. Said transgenic animals constitute a model for the study of said diseases and for the evaluation of compounds for the treatment and/or prevention of the diseases. Also disclosed are DNA construct and methods useful for producing the non-human transgenic mammals.

1 - 29 . (canceled)  
     
     
         30 . A polynucleotide construct that comprises a gene that is created or activated by a chromosomal anomaly associated with a human pathology of stem cell origin, wherein the gene is under control of a promoter that directs the expression of said gene in Sca-1 +  cells.  
     
     
         31 . A construct according to  claim 30 , wherein said chromosomal anomaly is a chromosomal anomaly associated with chronic myeloid leukaemia, B-cell acute lymphoblastic leukaemia, or T-cell acute lymphoblastic leukaemia, or with migration of haematopoietic or embryonic stem cells.  
     
     
         32 . A construct according to  claim 31 , wherein said gene is selected from the group consisting of BCR-ABL P210 , BCR-ABL p190 , Slug, Snail, HOX11, RHOM2/LMO-2 and TAL1.  
     
     
         33 . A construct according to  claim 32 , wherein said gene is selected from the group consisting of human BCR-ABL p210 , human BCR-ABL p190 , murine Slug, murine Snail, human HOX11, human RHOM2/LMO-2 and human TAL1 cells.  
     
     
         34 . A construct according to  claim 30 , wherein promoter that directs the expression in Sca-1 +  cells of said gene is a mouse promoter pLy-6E.1 or a functional fragment thereof.  
     
     
         35 . A transgenic non-human mammal that contains in its genome a DNA construct according to  claim 30 .  
     
     
         36 . A transgenic non-human mammal according to  claim 35 , wherein the mammal is a mouse.  
     
     
         37 . A transgenic non-human mammal according to  claim 36 , wherein the mammal displays a pathology of stem cell origin selected from the group of chronic myeloid leukaemia, B-cell acute lymphoblastic leukaemia, or T-cell acute lymphoblastic leukaemia, or a pathology associated with migration of haematopoietic or embryonic stem cells.  
     
     
         38 . A transgenic non-human mammal according to  claim 37 , wherein the transgenic mouse is selected from the group consisting of the mice identified as Sca-1 +  BCR-ABL p210 , Sca-1 +  BCR-ABL p190 , Sca-1 +  Slug, Sca-1 +  Snail, Sca-1 +  HOX11, Sca-1 +  RHOM2/LMO-2 and Sca-1 +  TAL1.  
     
     
         39 . A progeny of a transgenic non-human mammal according to  claim 35 .  
     
     
         40 . A cell line of a transgenic non-human mammal that contains in its genome a DNA construct according to  claim 30 .  
     
     
         41 . A cell line according to  claim 40 , wherein the cell line is a murine cell line.  
     
     
         42 . A method for producing a transgenic non-human mammal that possesses a chromosomal anomaly associated with a pathology of stem cell origin, the method comprising: 
 (i) introducing a DNA construct according to  claim 30  into a fertilized oocyte of a non-human transgenic mammal;    (ii) implanting said fertilised oocyte into a pseudopregnant wet nursing mother to produce a descendent; and    (iii) analysing said descendent to evaluate the presence of said created or activated by a chromosomal anomaly.    
     
     
         43 . A method according to  claim 42 , wherein the chromosomal anomaly associated with a human pathology of stem cell origin is a chromosomal anomaly associated with chronic myeloid leukaemia, B-cell acute lymphoblastic leukaemia, or T-cell acute lymphoblastic leukaemia, or with the migration of haematopoietic or embryonic stem cells.  
     
     
         44 . A procedure according to  claim 42 , wherein the non-human mammal is a mouse.  
     
     
         45 . A method for evaluating a candidate compound for treating or preventing a chromosomal anomaly associated with neoplastic or non-neoplastic human pathology of either haematopoietic or non-haematopoietic stem cell origin, the method comprising 
 administering a suitable amount of the candidate compound to a non-human transgenic mammal of  claim 37 , and    evaluating the effects of said compound on the occurrence or symptoms of said chromosomal anomaly.    
     
     
         46 . A method for evaluating a candidate compound for treating or preventing a chromosomal anomaly associated with neoplastic or non-neoplastic human pathology of either haematopoietic or non-haematopoietic stem cell origin, the method comprising 
 administering a suitable amount of the candidate compound to a cell of  claim 40 , and    evaluating the effects of said compound on the occurrence or symptoms of said chromosomal anomaly.    
     
     
         47 . A method according to  claim 44 , wherein the chromosomal anomaly associated with a human pathology of stem cell origin is a chromosomal anomaly associated with chronic myeloid leukaemia, B-cell acute lymphoblastic leukaemia, or T-cell acute lymphoblastic leukaemia, or with the migration of haematopoietic or embryonic stem calls.  
     
     
         48 . A method for treating or preventing a chromosomal anomaly associated with a human pathology of stem cell origin, the method comprising directing the expression of a suitable gene in Sca-1 +  cells using a promoter.  
     
     
         49 . A method according to  claim 48 , wherein the chromosomal anomaly associated with a human pathology of stem cell origin is a chromosomal anomaly associated with chronic myeloid leukaemia, B-cell acute lymphoblastic leukaemia, or T-cell acute lymphoblastic leukaemia, or with the migration of haematopoietic or embryonic stem calls.