《干细胞癌症描述课件.ppt》由会员分享，可在线阅读，更多相关《干细胞癌症描述课件.ppt（76页珍藏版）》请在三一办公上搜索。
1、Flatworm (planarian),Newt,MRL mice,Nature. 2012 Sep 27;489(7417):561-5. doi: 10.1038/nature11499. Skin shedding and tissue regeneration in African spiny mice (Acomys).Seifert AW, Kiama SG, Seifert MG, Goheen JR, Palmer TM, Maden M.,Flatworm (planarian)NewtMRL mi,Stem Cell Development,Adult stem/prog
2、enitors,functional maturation,PCD (apoptosis),Differentiation,Death,Senescence,ESCs,chronological aging,Stem Cell DevelopmentAdult ste,Stem Cells & Cancer,Three tumor biology puzzles:Most tumors are of a clonal origin but tumor cells are heterogeneous.It is very difficult to establish stable tumor c
3、ell lines from tumors.Large numbers of established tumor cells have to be injected to re-initiate an orthotopic tumor in mice.Key reviews:Reya T et al. Stem cells, cancer, and cancer stem cells. Nature 414, 105-111, 2001.Dick JE. Stem cell concepts renew cancer research. Blood 112: 4793-4807, 2008.V
4、isvader JE, and Lindeman GJ. Cancer Stem Cells: Current Status and Evolving Complexicities. Cell Stem Cell 10: 717-728, 2012.4. Tang DG. Understanding cancer stem cell heterogeneity and plasticity. Cell Res, 22(3):457-472, 2012. 5. Magee JA, Piskounova E, & Morrison SJ. Cancer Stem Cells: Impact, He
5、terogeneity, and Uncertainty. Cancer Cell 21: 283-296, 2012.(Dean Tang, Basic Concepts of Tumor Biology, Oct 31, 2012),Stem Cells & CancerThree tumor,1. Characteristics & Definition2. SC Identification3. SC Niche & Plasticity4. SCs & Cancer5. Cancer Stem Cells (CSCs),Stem Cells & Cancer,1. Character
6、istics & Definitio,Rare Generally small- Normally localized in a protected environment called NICHE, where they only occasionally divide. - But they possess HIGH PROLIFERATIVE POTENTIAL and can give rise to large clones of progeny in vitro or in vivo following injury or appropriate stimulation.- Pos
7、sess the ability to SELF-RENEW (i.e., asymmetric or symmetric cell division)- Can generate (i.e., DIFFERENTIATE into) one or multiple or all cell types (uni-, oligo-, multi-, pluri-, or toti-potent).,Stem Cells,Stem Cells,Committed progenitor cells,symmetric SC renewal,asymmetric celldivision (ACD),
8、symmetric SC commitment (differentiation),SC,Tang, Cell Res. 2012,Committed progenitor cellssymm,LT-SC,ST-SC,Lateprogenitors,Differen-tiated cells,Differen-tiating cells,Earlyprogenitors,Proliferation,Differentiation,Transformationprobability,Self-renewal,Niche,Commitment,Differentiation,Expansion,T
9、ang, Cell Res. 2012,Cell lineage development: Self-renewal, proliferation, & differentiation,LT-SCST-SCLateDifferen-Differe,Mouse ESCs were generated early 1980s by Evans and Martin. mES cells are cultured on mouse fibroblast feeders (irradiated or mitomycin C-treated) together with LIF.mES cells ar
10、e widely used in gene targeting.Human ES (hES) cells were first created by Jim Thomson (Uni. Wisconsin) in 1998. hES cells were initially cultured also on mouse fibroblast feeders but without LIF. Now they can be maintained in defined medium with high bFGF (100 ng/ml), activin, and some other factor
11、s.,Embryonic Stem Cells (ESCs),Embryonic Stem Cells (ESCs),Leftover or dead-end IVF embryos (PGD),How can hES cells be derived?,How can hES cells be derived?,16-cell morula,16-cell morula,Primitive ectoderm,Trophectoderm,Primitive Endoderm,A. Nagy,Primitive ectodermTrophectoder,ES cells,A. Nagy,ES c
12、ellsA. Nagy,TS cells,A. Nagy,TS cellsA. Nagy,A. Nagy,A. Nagy,A. Nagy,A. Nagy,A. Nagy,A. Nagy,heart,pancreas,testis,liver,brain,kidney,A. Nagy,heartpancreastestisliverbraink,干细胞癌症描述课件,Derived from umbilical cord Primarily blood stem cellsAlso contain mesenchymal stem cells that can differentiate into
13、 bone, cartilage, heart muscle, brain, liver tissue etc.*CB-SC could be stimulated to differentiate into neuron, endothelial cell, and insulin-producing cells,Cord Blood Stem Cells (CB-SC),Germline Stem Cells (GSC),Other embryonic SCs,Cord Blood Stem Cells (CB-SC)G,1. Characteristics & Definition2.
14、SC Identification3. SC Niche & Plasticity4. SCs & Cancer5. Cancer Stem Cells (CSCs),Stem Cells & Cancer,1. Characteristics & Definitio,Functional Assays of Stem Cells,(Candidate) Stem Cells,Stem Cells in situ,(Xeno)Transplantation,Lineage tracing,Functional Assays of Stem Cell,How to identify and ch
15、aracterize (adult) stem cells?,Marker analysisLabel-retaining cells (LRC): Pulse-chase exper.Clonal/clonogenic assaysFunctional analysis: Side population (SP) assayFunctional analysis: Aldefluor assayCell size-based enrichmentGenetic marking & lineage tracing,How to identify and characteri,Passegu,
16、Emmanuelle et al. (2003) Proc. Natl. Acad. Sci. USA 100, 11842-11849,Hematopoietic stem/progenitor cell lineages,(1:5,000 or 0.02%;lifetime self-renewal),(1:1,000 or 0.1%;self-renewal for 8 wks),(No self-renewal),Lin-Sca1+ckit+CD150+CD48- (20%-50% such mouse BM cells are SCs),Till JE & McCulloch EA.
17、 A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. Radiat. Res 14, 213-222, 1961.,Lin-CD34+CD38-CD45RA-Thy1+RholoCD49f+(Notta F.Dick JE. Science 333, 218-221, 2011),Passegu, Emmanuelle et al. (2,(Nestin),(GFAP),(Pax6),(A2B5),(NG2),(MBP),(NeuM),(Mash-1),(PDGFRa),(Ne
18、stin)(GFAP)(Pax6)(A2B5)(NG2,Sue Fischer,Sue Fischer,How to identify and characterize (adult) stem cells?,Marker analysisLabel-retaining cells (LRC): Pulse-chase exper.Clonal/clonogenic assaysFunctional analysis: Side population (SP) assayFunctional analysis: Aldefluor assayCell size-based enrichment
19、Genetic marking & lineage tracing,How to identify and characteri,Till JE & McCulloch EA. A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. Radiat. Res 14, 213-222, 1961. The earliest report in which putative stem cells were identified by their ability to retain lab
20、eled radionucleotides for long period of timeCotsarelis G, Cheng SZ, Dong G, Sun TT & Lavker RM. Existence of slow-cycling libmal epithelial basal cells that can be preferentially stimulated to proliferate: Implications on epithelial stem cells. Cell 57, 201-209, 1989.Cotsarelis G, Sun TT, & Lavker
21、RM. Label-retaining cell reside in the bulge area of pilosebaceous unit: implications for follicular stem cells, hair cycle, and skin carcinogenesis. Cell 61, 1329-1337, 1990.,Till JE & McCulloch EA. A dire,LRCs in the Bulge & BM ARE Stem Cells,Tumbar T et al., Defining the epithelial stem cell nich
22、e in skin. Science 303, 359-363, 2004. Blanpain, C., et al., Self-renewal, multipotency, and the existence of two cell populations in an epithelial stem cell niche. Cell 118, 635-648, 2004.Fuchs et al., Cell 116, 769, 2004Fuchs E: The tortoise and the hair: Slow-cycling cells in the stem cell race.
23、Cell 137, 811-819, 2009.Fuchs E & Horsley V. Ferreting out stem cells from their niches. Nat Cell Biology 13: 513-518, 2011.Wilson A et al. Hematopoietic stem cells reversibly switch from dormancy to self-renewal during homeostasis and repair. Cell 135, 1118-1129, 2008.Foudi A et al. Analysis of his
24、tone 2B-GFP retention reveals slowly cycling hematopoietic stem cells. Nat. Biotechnol.27, 84-90, 2009.,LRCs in the Bulge & BM ARE Ste,Not All Stem Cells Are Slow-Cycling,Fuchs E & Horsley V. Ferreting out stem cells from their niches. Nat Cell Biology 13: 513-518, 2011.Li L & Clevers H. Co-existenc
25、e of quiescent and active adult stem cells in mammals. 327, 542-545, 2010.,. and even for the ones that do, approximately only 5-6 divisions of the label-retaining stem cell or its progeny can be monitored after a pulse-chase beforethe label has diluted out to the point where it can no longer be tra
26、ced.,Not All Slow-Cycling Cells Are Stem Cells,Not All Stem Cells Are Slow-Cy,How to identify and characterize (adult) stem cells?,Marker analysisLabel-retaining cells (LRC): Pulse-chase exper.Clonal/clonogenic assaysFunctional analysis: Side population (SP) assaysFunctional analysis: Aldefluor assa
27、yCell size-based enrichmentGenetic marking & lineage tracing,How to identify and characteri,E,Rheinwald JG & Green H. Serial cultivation of human epidermal keratinocytes: The formation of keratinizing colonies from single cells. Cell 6, 331-343, 1975.,Sun TTCell 9, 511-521, 1976Nature 269, 489-493,
28、1977Cell 14, 469-476, 1978,Fuchs ECell 19, 1033-1042, 1980Cell 25, 617-625, 1981,Barrandon YPNAS 82, 5390-4, 1985Cell 50, 1131-1137, 1987,Rice RHCell 11, 417-422, 1977Cell 18, 681-694, 1979,Watt FMJCB 90, 738-742, 1981,ERheinwald JG & Green H. Seri,CLONAL vs CLONOGENIC ASSAYS,Cloning efficiency (CE;
29、 %)Clonal size (cell number/clone)Clonal development (tracking)Clone types,A clone: a two-dimensional structure,CLONAL vs CLONOGENIC ASSAYS,Mixing Experiments to Demonstrate the Clonality of Clones/Spheres,DU145 RFP:DU145 GFP (1:1) MC,DU145:DU145 GFP (1:1) MC,phase,GFP,DU145:DU145 GFP (1:1) Clonal A
30、ssay,Pastrana E, Silva-Vargas V, and Doetsch F.Eyes wide open: A critical review of sphere-formation as an assay of stem cells.Cell Stem Cell 8, 486-498, 2011,Mixing Experiments to Demonst,How to identify and characterize (adult) stem cells?,Marker analysisLabel-retaining cells (LRC): Pulse-chase ex
31、per.Clonal/clonogenic assaysFunctional analysis: Side population (SP) assaysFunctional analysis: Aldefluor assayCell size-based enrichmentGenetic marking & lineage tracing,How to identify and characteri,Goodell MA et al. Isolation and functional properties of murine hematopoietic stem cells that are
32、 replicating in vivo. J. Exp. Med. 183, 1797-1806, 1996.,Golebiewska A, Brons NH, Bjerkvig R, and Niclou SP. Critical appraisal of the side population assayin stem cell and cancer stem cell research. Cell Stem Cell 8, 136-147, 2011.,Goodell MA et al. Isolation an,Bcrp (ABCG2) is a major mediator of
33、the SP phenotype,Zhou et al., Nature Med 7, 1028, 2001,Bcrp (ABCG2) is a major mediat,How to identify and characterize (adult) stem cells?,Marker analysisLabel-retaining cells (LRC): Pulse-chase exper.Clonal/clonogenic assaysFunctional analysis: Side population (SP) assaysFunctional analysis: Aldefl
34、uor assayCell size-based enrichmentGenetic marking & lineage tracing,How to identify and characteri,Alison MR, Guppy NJ, Lim SML, & Nicholson LJ. Finding cancer stem cells: Are aldehyde dehydrogenases fitfor purpose? J Pathol. 222, 335-344, 2010.Ma I & Allan AL. The role of human aldehyde dehydrogen
35、ase in normal and cancer stem cells. Stem Cell Rev and Report 7, 292-306, 2011.,*ALDH1A1 and ALDH3A1: Seem to be the major isozymes mediating the Aldefluor phenotype and are preferentially expressed in SC.*ALDH superfamily: 19 putatively functional genes in 11 families and 4 subfamilies. ALDH superf
36、amily of NAD(P)+-dependent enzyme catalyzes oxidations of aldehydes to carboxylic acids.,Kastan MB et al. Direct demonstration of elevated aldehyde dehydrogenase in human hematopoieticprogenitor cells. Blood 75, 1947-1960, 1990.Jones RJ et al., Assessment of aldehyde dehydrogenase in viable cells. B
37、lood 85, 2742-46, 1995.Storms RW et al. Isolation of primitive human hematopoietic progenitors on the basis of aldehydedehydrogenase activity. PNAS 96, 9118-9123, 1999.,Alison MR, Guppy NJ, Lim SML,How to identify and characterize (adult) stem cells?,Marker analysisLabel-retaining cells (LRC): Pulse
38、-chase exper.Clonal/clonogenic assaysFunctional analysis: Side population (SP) assaysFunctional analysis: Aldefluor assayCell size-based enrichmentGenetic marking & lineage tracing,How to identify and characteri,Fuchs E & Horsley V. Ferreting out stem cells from their niches. Nature Cell Biology 13:
39、 513-518, 2011.,Fuchs E & Horsley V. Ferreting,Fuchs E & Horsley V. Ferreting out stem cells from their niches. Nature Cell Biology 13: 513-518, 2011.Liver J et al., Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system.Nature 450, 56-62, 2007.Snippert HJ e
40、t al., Intestinal crypt homeostasis results from neutral competition between symmetrically dividingLrg stem cells. Cell 143, 134-144, 2010.,Fuchs E & Horsley V. Ferreting,1. Characteristics & Definition2. SC Identification3. SC Niche & Plasticity4. SCs & Cancer5. Cancer Stem Cells (CSCs),Stem Cells
41、& Cancer,1. Characteristics & Definitio,Stem Cell Niche in Hair Follicles: The Bulge,Moore KA & Lemischka IR. Science 311, 1880-1885, 2006,Stem Cell Niche in Hair Follic,Bulge Stem Cells,Tumbar et al., Science 303, 359-363, 2004; Fuchs et al., Cell 116, 769, 2004,Bulge Stem CellsTumbar et al.,Stem C
42、ell Niche in Small Intestine: The Crypt,Moore KA & Lemischka IR. Science 311, 1880-1885, 2006,Barker N et al., Cell Stem Cell 11: 452-460, 2012.,Stem Cell Niche in Small Intes,Stem Cell Niches in BM,Moore KA & Lemischka IR. Science 311, 1880-1885, 2006,Naveiras O et al., Bone-marrow adipocytes as ne
43、gative regulators of the hematopoietic microenvironment. Nature 460, 259, 2009.Mendez-Ferrer, S et al., Mesenchymal and hematopoietic stem cells form a unique bone marrow niche. Nature 466, 829-834, 2010.,Stem Cell Niches in BMMoore KA,Stem cell lineage,Differentiatedcells,Death (PCD),Senescence,Ste
44、m cells,Progenitors/Precursor cells,Other differ.cell(s),Stem cell lineageDifferentiate,*First report: Long-term cultured adult brain (stem) cells can reconstitute the whole blood in lethally irradiated mice (Bjornson et al., Science 283, 534-537, 1999).*Cells from skeletal muscle have hematopoietic
45、 potential (Jackson et al., PNAS 96, 14482-14486, 1999) and can also “differentiate” into many other cell types (Qu-Petersen, Z, et al., JCB 157, 851-864, 2002).*CNS “SCs” can “differentiate” into muscle cells (Clarke et al., Science 288, 1660-1663, 2000; Galli et al., Nat. Neurosci 3, 986-991, 2000
46、; Tsai and McKay, J. Neurosci 20, 3725-3735, 2000).*Vice versa, “SCs” from blood or bone marrow can “transdifferentiate” into muscle (Ferrari et al., Science 279, 1528-1530, 1998; Gussoni et al., Nature 401, 390-394, 1999), hepatocytes (Petersen et al., Science 284, 1168-1170, 1999; Lagasse et al.,
47、Nat Med 6, 1229-1234, 2000), cardiac myocytes (Orlic et al., Nature 410, 701-705, 2001), or neural cells (Mezey et al., Science 290, 1779-1782, 2000; Brazelton et al., Science 290, 1775-1779, 2000).*Bone marrow appears to contain two distinct SCs: the HSC and MSC. A single HSC could contribute to ep
48、ithelia of multiple organs of endodermal and ectodermal origin (Krause et al., Cell 105 369-377, 2001). MSC, on the other hand, can adopt a wider range of fates (endothelial, liver, neural cells, and perhaps all cell types) (Pittenger et al., Science 284, 143-146, 1999; Schwartz et al., JCI 109, 129
49、1-1302, 2002; Jiang et al., Nature 418, 41-49, 2002).*Pluripotent “SCs” have also been isolated from skin that can “differentiate” into neural cells, epithelial cells, and blood cells (Toma et al., Nat Cell Biol. 3, 778-784, 2001)*Highly purified adult rat hepatic oval “stem cells, which are capable
50、 of differentiating into hepatocytes and bile duct epithelium, can “trans-differentiate” into pancreatic endocrine hormone-producing cells when cultured in a high glucose environment (Yang et al., PNAS 99, 8078-8083, 2002),“Transdifferentiation” of Stem Cells,*First report: Long-term cultu,De-differ
