Humanized mouse model with improved human innate immune cell development

Abstract

A genetically-modified, immunodeficient mouse is provided along with methods of use, wherein the mouse includes (a) a nucleotide sequence encoding human stem cell factor (hSCF); (b) a nucleotide sequence encoding human granulocyte-macrophage colony-stimulating factor (hGM-CSF); (c) a nucleotide sequence encoding human interleukin-3 (hIL-3); and (d) a nucleotide sequence encoding human colony-stimulating factor 1 (hCSF1), wherein each of the nucleotide sequences is operably linked to a promoter, and wherein the genetically-modified, immunodeficient mouse expresses hSCF, hGM-CSF, hIL-3, and hCSF1, wherein the genetically-modified, immunodeficient mouse allows engraftment of human hematopoietic stem cells along with engraftment of human-patient derived tumor xenografts and/or human tumor cell lines to enable in vivo investigation of the interactions between the human immune system and human cancer.

1 . A genetically-modified, immunodeficient mouse that expresses human stem cell factor (hSCF), human granulocyte-macrophage colony-stimulating factor (hGM-CSF), human interleukin-3 (hIL-3), and human colony-stimulating factor 1 (hCSF1). 
     
     
         2 . The genetically-modified, immunodeficient mouse of  claim 1 , wherein the mouse is a NOD.Cg-Prkdc scid Il2rg tm1Wjl /SzJ (NSG) mouse, a NOD.Cg-Rag1 tm1Mom  Il2rg tm1Wjl /SzJ (NRG) mouse, or NOD.Cg-Prkdc scid  Il2rg tm1Sug /JicTac (NOG) mouse. 
     
     
         3 . (canceled) 
     
     
         4 . The genetically-modified, immunodeficient mouse of  claim 1 , further comprising a human hematopoietic stem cell. 
     
     
         5 . The genetically-modified, immunodeficient mouse of  claim 1 , wherein the mouse comprises a differentiated human hematopoietic stem cell selected from the group consisting of a human myeloid progenitor cell, a human lymphoid progenitor cell, a human CD33 +  myeloid cell, a human mast cell, a human monocyte, a human macrophage, human myeloid dendritic cell, a human B cell, a human plasma cell, a human T cell, a human T helper cell, a human cytotoxic T cell, a human T reg  cell, and a human natural killer cell. 
     
     
         6 . The genetically-modified, immunodeficient mouse of  claim 1 , wherein the mouse comprises a human leukocyte selected from the group consisting of a human CD45 + , CD20 + , CD20+CD45 + , CD3 + , CD3 + CD45 + , CD33 + , CD33 + CD45 + , CD14 + , CD14 + CD45 + , CD56 + , and CD56 + CD45 +  leukocyte. 
     
     
         7 . The genetically-modified, immunodeficient mouse of  claim 6 , wherein the mouse comprises, in the absence of an immunological challenge:
 at least about 20% of the human CD45 +  leukocytes of the mouse are CD3 + CD45 +  leukocytes;   at least about 10% of the human CD45 +  leukocytes of the mouse are CD33 + CD45 +  leukocytes;   at least about 5% of the human CD45 +  leukocytes of the mouse are CD14 + CD45 +  leukocytes; or   at least about 0.5% of the human CD45 +  leukocytes of the mouse are CD56 + CD45 +  leukocytes.   
     
     
         8 . The genetically-modified, immunodeficient mouse of  claim 4 , wherein the mouse expresses a human cytokine selected from the group consisting of human interleukin-8, human interleukin-1β, human tumor-necrosis factor, human interleukin-12p70, and human interleukin-6. 
     
     
         9 . The genetically-modified, immunodeficient mouse of  claim 1 , further comprising a human xenograft comprising a human tumor cell. 
     
     
         10 . A method of making a genetically-modified, immunodeficient humanized mouse model, comprising
 administering human hematopoietic stem cells to the genetically-modified, immunodeficient mouse of  claim 1 .   
     
     
         11 . The method of  claim 10 , further comprising conditioning the genetically-modified, immunodeficient mouse to reduce mouse hematopoietic cells of the mouse prior to administering the human hematopoietic stem cells. 
     
     
         12 . The method of  claim 11 , wherein the conditioning comprises irradiating the genetically-modified, immunodeficient mouse and/or administering a radiomimetic drug to the genetically-modified, immunodeficient mouse. 
     
     
         13 . The method of  claim 10 , further comprising administering a human xenograft comprising a human tumor cell to the genetically-modified, immunodeficient mouse. 
     
     
         14 . The method of  claim 13 , wherein the human hematopoietic stem cell and the human tumor cell comprise at least 2 matching HLA alleles. 
     
     
         15 . A method of identifying anti-tumor activity of a test substance, comprising:
 administering a human tumor cell to the genetically-modified, immunodeficient mouse of  claim 1 , wherein the human tumor cell form a solid or non-solid tumor in the genetically-modified, immunodeficient mouse;   administering a test substance to the genetically-modified, immunodeficient mouse; and   assaying a response of the solid or non-solid tumor to the test substance, wherein an inhibitory effect of the test substance on the tumor and/or tumor cells identifies the test substance as having anti-tumor activity.   
     
     
         16 . The method of  claim 15 , further comprising comparing the response to a standard to determine the effect of the test substance on the xenogeneic tumor cell, wherein an inhibitory effect of the test substance on the xenogeneic tumor cell identifies the test substance as having anti-tumor activity. 
     
     
         17 . The method of  claim 15 , wherein the test substance is an immunotherapeutic agent. 
     
     
         18 . The method of  claim 15 , wherein the test substance is an immune checkpoint inhibitor. 
     
     
         19 . (canceled) 
     
     
         20 . (canceled) 
     
     
         21 . The method of  claim 15 , wherein the test substance is an antibody. 
     
     
         22 . The method of  claim 15 , wherein the test substance is an anti-cancer agent. 
     
     
         23 .- 25 . (canceled) 
     
     
         26 . The genetically-modified, immunodeficient mouse of  claim 1 , wherein the mouse comprises a nucleotide sequence encoding the hSCF, a nucleotide sequence encoding the hGM-CSF, a nucleotide sequence encoding the hIL-3, and a nucleotide sequence encoding the hCSF1, wherein each of the nucleotide sequences is operably linked to a promoter.