Genetically modified non-human animals expressing human EPO from the animal genome are provided. Also provided are methods for making non-human animals expressing human EPO from the non-human animal genome, and methods for using non-human animals expressing human EPO from the non-human animal genome. These animals and methods find many uses in the art, including, for example, in modeling human erythropoiesis and erythrocyte function; in modeling human pathogen infection of erythrocytes; in in vivo screens for agents that modulate erythropoiesis and/or erythrocyte function, e.g. in a healthy or a diseased state; in in vivo screens for agents that are toxic to erythrocytes or erythrocyte progenitors; in in vivo screens for agents that prevent against, mitigate, or reverse the toxic effects of toxic agents on erythrocytes or erythrocyte progenitors; in in vivo screens of erythrocytes or erythrocyte progenitors from an individual to predict the responsiveness of an individual to a disease therapy.

A mouse with a humanization of the miL-3 gene and the mGM-CSF gene, a knockout of a mRAG gene, and a knockout of a mII2rg subunit gene; and optionally a humanization of the TPO gene is described. A RAG/II2rg KO/hTPO knock-in mouse is described. A mouse engrafted with human hematopoietic stem cells (HSCs) that maintains a human immune cell (HIC) population derived from the HSCs and that is infectable by a human pathogen, e.g., S. typhi or M. tuberculosis is described. A mouse that models a human pathogen infection that is poorly modeled in mice is described, e.g., a mouse that models a human mycobacterial infection, wherein the mouse develops one or more granulomas comprising human immune cells. A mouse that comprises a human hematopoietic malignancy that originates from an early human hematopoietic cells is described, e.g., a myeloid leukemia or a myeloproliferative neoplasia.

A recombinant protein is provided. The recombinant protein of the invention comprises an erythropoietin and a highly glycosylated peptide, and has a longer half-life. Further, the recombinant protein of the invention may also comprise a carboxyl-terminal peptide of human chorionic gonadotropin and a carboxyl-terminal peptide of thrombopoietin.

Bone-targeting therapeutic compounds and pharmaceutically acceptable salts thereof; pharmaceutical compositions comprising a bone-targeting therapeutic compound; processes for preparing bone-targeting therapeutic compounds; and therapeutic methods to treat bone defects.

A method for treating a defect site in a living bone of an animal by applying an exogenous compound having thrombopoietic activity to the defect site in an amount effective to induce thrombopoiesis. The exogenous compound activates a thrombopoietin receptor, leading to accelerated bone formation at the defect site. Also provided is a method for repairing a segmental bone defect in an animal bone by inserting into the segmental bone defect a biodegradable bone repair scaffold that contains a compound having thrombopoietic activity. The compound activates a thrombopoietin receptor and accelerates bone formation such that bridging occurs at the segmental bone defect.

TPO was used to promote the growth of bone in both rats and in mice. Gaps in both mouse and in rat bones were treated with a scaffold sized to fit the gap. Scaffolds that included TPO promoted better outcomes than scaffolds that included BMP-2 or scaffolds that did not include either TPO or BMP-2. These data indicate that compounds that exhibit thrombopoietic activity such a recombinant TPO can be used to promote bone growth and healing in mammals.

The invention relates generally to genetically modified non-human animals expressing human polypeptides and their methods of use.

This invention concerns a process for making a lyophilized pharmaceutical formulation of a therapeutic protein, which comprises (a) providing a formulation of a bulk amount of the therapeutic protein, (b) measuring the concentration of the therapeutic protein in said bulk formulation, (c) adjusting the fill weight of the protein in said bulk formulation to achieve a fixed dose of the protein, and (d) lyophilizing the protein fill weight-adjusted formulation to achieve a final formulation in a container, wherein the product concentration post reconstitution with a fixed volume is within a predetermined acceptance range. The process is particularly suitable for formulations with low protein concentrations (e.g., 0.05 to 20 mg/mL).

Genetically modified non-human animals are provided that may be used to model human hematopoietic cell development, function, or disease. The genetically modified non-human animals comprise a nucleic acid encoding human IL-6 operably linked to an IL-6 promoter. In some instances, the genetically modified non-human animal expressing human IL-6 also expresses at least one of human M-CSF, human IL-3, human GM-CSF, human SIRPa or human TPO. In some instances, the genetically modified non-human animal is immunodeficient. In some such instances, the genetically modified non-human animal is engrafted with healthy or diseased human hematopoietic cells. Also provided are methods for using the subject genetically modified non-human animals in modeling human hematopoietic cell development, function, and/or disease, as well as reagents and kits thereof that find use in making the subject genetically modified non-human animals and/or practicing the subject methods.

The recombinant peptides and proteins having thrombopoietin (TPO) activities, e.g., thrombopoietin mimetic peptides that are TPO receptor (TPOR) agonists. The recombinant peptides or proteins include a recombinant human p75 TNF receptor and TPO mimetic peptides linked by an Fc fragment of IgG1. The recombinant peptides form a homodimer by inter-chain disulfide bond in the Fc region and include six TPOR binding and/or activating domain in the C-terminus. The recombinant peptides or proteins provided herein are useful for increasing platelet count, e.g., for treating thrombocytopenia. The methods for producing the recombinant peptides and proteins, formulations, and therapeutic methods.