Microsoft PowerPoint - 11 Hematopoietic Growth Factor.pptx

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1 Pharmaceutical Biotechnology Hematopoietic Growth Factor By Yuqiong Xia 2014

2 Keywords Hematopoietic growth factor (HGF), hematopoietic stem cells (HSC) Red blood cell, white blood cell, lymphocyte, platelet, megakaryocyte, macrophagecyte G-CSF, GM-CSF, EPO, TPO, MGDF, SCF 2

3 Outline Stem cells and hematopoietic stem cells Hematopoiesis Hematopoietic growth factors 3

4 Stem cell 生物制药工程夏玉琼西安电子科技大学 4

5 Embryonic stem cell 5

cell Epithelial cell Fat cell Neuron 6 Cardiac muscle

6 Adult stem cell Liver Bone Brain CNS stem cell Bone marrow Skeletol muscle Blood cell Blood vessel Bone marrow stomal cell Epithelial cell Fat cell Neuron 6 Cardiac muscle

7 Hematopoietic stem cells Hematopoietic stem cells (HSCs) are the blood cells that give rise to all the other blood cells and are derived from mesoderm( 中胚层 ) HSCs are found in the bone marrow of adults, with large quantities in the pelvis, femur, and sternum( 盆骨股骨和胸骨 ). They are also found in umbilical cord blood ( 脐带血 )and, in small numbers, in peripheral blood( 外周血 ). 7

8 How to get HSC from bone marrow Stem and progenitor cells can be taken from the pelvis, at the iliac crest, using a needle and syringe. 髂骨 8

9 How to get HSC from peripheral blood Blood donors are injected with a cytokine, such as granulocyte-colony stimulating factor (G-CSF) The cytokine induce cells to leave the bone marrow and circulate in the blood vessels Draw the blood 9

Bone marrow and its cells 正常红细胞 normoblast with dividing nucleus 髓细胞 myelocyte 红细胞 erythrocyte 破骨细胞 myeloplaxe 嗜酸性粒细胞 Eosinophil cell Normoblast

10 Bone marrow and its cells 正常红细胞 normoblast with dividing nucleus 髓细胞 myelocyte 红细胞 erythrocyte 破骨细胞 myeloplaxe 嗜酸性粒细胞 Eosinophil cell Normoblast Myelocyte dividing Myelocyte Myelocyte Fat Fat 10

11 The constitution of blood 11

Red blood cell Also called erythrocyte D = 6.2-8.

12 Red blood cell Also called erythrocyte D = micrometers Lack cell nucleus Transport oxygen and carbon dioxide 12

13 White blood cell (WBC) Also called leukocytes Larger than RBC 7000 WBC per microliter blood ~1% of the blood volume Defend the body against both infectious diseases and foreign materials 13

14 Classifications of WBC Granulocytes 粒细胞 Neutrophil (most) Eosinophil (big particles) Basophil (least, < 0.6%) Agranulocytes 无颗粒的白细胞 Lymphocyte Monocyte (biggest WBC) 14

15 RBC vs. WBC RBC WBC Physical feature No nucleus, bioconcave disc Nucleus with outer white buffer coat, irregular shape shaped Life span 120 days 4-30 days Type One Various Circulatory systems Counts in blood Function Cardiovascular systems Cardiovascular and lymphatic systems 36%-50% of blood 1% of blood Supply O 2 and transport CO Producing antibodies to develop immunity against infections

Platlet Platlet blood bag 16 http://simple.

16 Platelet Also called thrombocytes D = 2-3 micrometers Lead to formation of blood clot Platlet Platlet blood bag 16

17 The origin of platlet: megakaryocyte One megakaryocyte ( 巨核细胞 ) can produce 2000 platlet on average Megakaryocyte is the largest cell found in red bone marrow 17

18 Outline Stem cells and hematopoietic stem cells Hematopoiesis Hematopoietic growth factors 18

19 The process of hematopoiesis Unsymmetrical division 生物制药工程 Lymphocyte夏玉琼西安电子科技大学淋巴细胞红细胞血小板巨噬细胞定向祖细胞嗜碱性细胞嗜酸性细胞嗜中性细胞 19 Megakaryocyte 巨核细胞 /CH17%20Hematopoiesis.htm

20 hematopoiesis = hemopoiesis ( 造血作用 ) The formation of blood Mediated by a series of growth factors The growth factors act individually and in various combinations stimulate the proliferation, differentiation, and function of hematopoietic cells 20

21 Outline Stem cells and hematopoietic stem cells Hematopoiesis Hematopoietic growth factors Function, composition and production Classification 21

22 What is hemopoietic growth factor (HGF) Regulatory proteins and local hormones Regulate the differentiation and proliferation of particular progenitor cells 22

23 What is the chemical composition of HGF Glycoproteins Amino acids Glycosylation Recombinant HGFs do not have carbohydrates HGFs are mostly single polypeptides 23

24 How to produce HGF Identify the gene in question and isolate it Insert the gene of interest into a plasmid, and then expressing the protein of interest in a biologic system (e.g., bacteria, yeast, or mammalian cells) to produce recombinant growth factors. 24

25 Outline Stem cells and hematopoietic stem cells Hematopoiesis Hematopoietic growth factors Function, composition and production Classification 25

26 Classification of HGFs The white cell factors The red cell factors The platelet factors Stem cell factor (SCF) 26

27 The white cell factors Granulocyte colony-stimulating factor (G-CSF) 粒细胞集落刺激因子 Granulocyte macrophage colony-stimulating factor (GM-CSF) 粒细胞巨噬细胞集落刺激因子 27

28 G-CSF Also known as colony-stimulating factor 3 (CSF 3) A glycoprotein that stimulates the bone marrow to produce granulocytes and stem cells and release them into the bloodstream G-CSF also stimulates the survival, proliferation, differentiation, and function of neutrophil precursors and mature neutrophils Produced by endothelium, macrophages, and a number of other immune cells 28

29 Therapeutic use of G-SCF To cancer patients To accelerate recovery from neutropenia after chemotherapy To HSC donor To increase the number of hematopoietic stem cells To HSC receiver To compensate for conditioning regimens. 29

30 GM-CSF GM-CSF is a protein secreted by macrophages, T cells, mast cells, NK cells, endothelial cells and fibroblasts. GM-CSF stimulates stem cells to produce granulocytes (neutrophils, eosinophils, and basophils) and monocytes. Monocytes exit the circulation and migrate into tissue, whereupon they mature into macrophages and dendritic cells. 30

31 Therapeutic use of GM-SCF To cancer patients Prevent neutropenia following chemotherapy To HIV-infected patients A vaccine adjuvant, but not FDA approved 31

32 G-CSF and GM-CSF as drugs G-CSF On chromosome AA GM-CSF On chromosome or 128 AA Filgrastim ( 非格司亭 ) rhg-csf (G-CSF produced using recombinant DNA technology) Marketed by several companies 32

33 The red cell factors Erythropoietins (EPO) 促红细胞生成素 darbepoetin α 阿法达贝泊汀 33

34 EPO 促红细胞生成素 Erythropoietin = EPO EPO is a glycoprotein hormone that stimulates the production of red blood cells by stem cells in the red bone marrow EPO is produced mainly by the kidneys EPO is released in response to decreased levels of oxygen detected in the blood 34

35 35

36 EPO In chromosome AA Require glycosylation for its in vivo biologic activity Two rhepo products are marketed and both are expressed in Chinese hamster ovary cells Epoetin α, in US, JAPAN, CHINA Epoetin β in US, JAPAN 36

37 Darbepoetin α 阿法达贝泊汀 Glycoengineered erythropoietin With more carbohydrate than rhepo With increased number of sialic acids ( 唾液酸 ) Sialic acids 37

38 The platelet factors Thrombopoietin (TPO) 促血小板生成素 Megakaryocyte growth and development factor (MGDF) 巨核细胞发育形成因子 Interleukin-11 (IL-11) 白介素 38

39 TPO A glycoprotein growth factor released by the liver, kidney and bone marrow in response to a reduction in circulating platelets Target the committed stem cells in the megakaryocyte lineage in red bone marrow to differentiate and proliferate Platelet production will increase to restore homeostasis 39

40 TPO On chromosome 3, with AA 40

41 MGDF MGDF is a truncated protein, with homology with the EPO-like amino-terminus of human TPO MGDF has identical biological properties to TPO in vitro and in vivo However, MGDF has a short circulatory half-life, which can be markedly increased when coupled to Clinical trials with PEG-rHuMGDF commenced in Australia and the United States in May

42 IL-11 白介素 11 IL-11 is a key regulator in stimulation of megakaryocyte maturation The human IL-11 gene, consisting of 5 exons and 4 introns, is located on chromosome 19, and encodes a 23 kda protein (199 AA) 42

43 Therapeutical use of IL-11 To cancer patients Improve platelet recovery after chemotherapyinduced thrombocytopenia Treat osteoporosis Participate in the regulation of bone cell proliferation and differentiation Oprelvekin (rhil-11) non-glycosylated polypeptide (177 AA) Marketed as Neumega in US 43

44 Stem cell factor (SCF) Fibronectin 纤连蛋白 Kit receptor SCF 44 Stromal cells

45 Stem cell factor (SCF) Increase the survival of HSCs in vitro and contributes to the self-renewal and maintenance of HSCs in-vivo SCF has been shown to increase adhesion and thus may play a large role in ensuring that HSCs remain in the niche A small percentage of HSCs regularly leave the bone marrow to enter circulation and then return to their niche in the bone marrow. SCF, along with the chemokine SDF-1, allow HSCs to find their way back to the niche. 45

46 Stem cell factor (SCF) An early acting HGF On chromosome 12 rhscf (ancestim), marketed as in Canada, Australia, and New Zealand 46

47 Summary of HGF 生物制药工程夏玉琼西安电子科技大学 47

48 Summary Stem cell, hematopoietic stem cell Hematopoiesis HGFs For white blood cell, G-CSF and GM-CSF For red blood cell, EPO For platelet, TPO, MGDF, IL-11 For stem cells, SCF 48

49 Homework 7 What are differences of G-CSF and GM-CSF about effects on cancer patients? What is the physiologic role of EPO? 49

Microsoft PowerPoint - 10 Hematopoietic Growth Factor.pptx

Microsoft PowerPoint - 10 Hematopoietic Growth Factor.pptx Pharmaceutical Biotechnology Hematopoietic Growth Factor By Yuqiong Xia 5/12/2017 生物技术制药 2 技术基础生物技术药物生物技术制药概念药物的开发药物的生产细胞因子生长因子激素类药物血液制品和治疗性酶抗体疫苗和佐剂核酸药物药物的发现专利的申请临床前试验临床试验生物药物的来源生物药物的生产和纯化生物药物的分析干扰素白介素造血生长因子