Bone Morphogenetic Proteins: Methods and Protocols

265 Pages·2018·6.68 MB·English

by  Melissa B. Rogers

Methods in
Molecular Biology 1891
Melissa B. Rogers Editor
Bone
Morphogenetic
Proteins
Methods and Protocols
M M B
ETHODS IN OLECULAR IO LO GY
SeriesEditor
JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
Hatfield, Hertfordshire AL109AB,UK
Forfurther volumes:
http://www.springer.com/series/7651
Bone Morphogenetic Proteins
Methods and Protocols
Edited by
Melissa B. Rogers
New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA
Editor
MelissaB.Rogers
NewJerseyMedicalSchool
RutgersBiomedicalandHealthSciences
Newark,NJ,USA
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-4939-8903-4 ISBN978-1-4939-8904-1 (eBook)
https://doi.org/10.1007/978-1-4939-8904-1
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Preface
The 31 members of the transforming growth factor β superfamily profoundly influence
animal biology from the earliest stages of germ cell development through life. Those
classifiedwithinthebonemorphogeneticprotein(BMP)brancharethemostevolutionarily
ancientmembers.Althoughfirstidentifiedbytheirosteogenicproperties,theseproteinsare
criticalsignalsinalldevelopingandadultinvertebrateandvertebratetissues.BMPorthologs
from distant species suchas flies and humans are functionally interchangeable and play key
roles in fundamental metazoan processes such as dorso-ventral axis determination,
epithelial-mesenchymal interactions, and programmed cell death to name just a few. Dis-
ruptions of BMP regulation and signaling cause or influence the entire panoply of human
clinicalconditions—cancers,cardiovascularandlungdiseases,osteogenicdiseasesinvolving
thin oroverly densebones and pathological soft tissue calcification,etc.Consequently,the
hopeofdiscoveringnewclinicalstrategiesforbothcommondiseasessuchasatherosclerosis
andrarediseasessuchasfibrodysplasiaossificansprogressiva(FOP)drivesvigorousresearch
intothebiologyofBMPs.
ThisvolumereviewsmethodstostudyBMPsincellcultureandinbothinvertebrateand
vertebrateanimals,andtherapeuticapplicationsofBMPsandBMPmodulators.Thetopics
havebeenchosentoextendandcomplementtherecentMethodsinMolecularBiologyvolume
entitled TGF-β Signaling [1]. Specifically, a greater focus has been placed on genetic
approaches and model organisms including both invertebrates and nonmammalian
vertebrates.
The book begins with a cell and molecular focus before moving into invertebrate
models and then ending with vertebrate models. The first three chapters discuss primary
andimmortalizedcelllinesusedforinvitrodifferentiationstudiesaswellasthedeliveryof
BMPs for cell-based gene therapy. Cutting-edge methodologies addressing transcription
and receptor mechanisms using genome editing and high-throughput biosensors are then
presented. The powerful genetic approaches to understanding BMP function in Caenor-
habditiselegansandDrosophilamelanogasterarepresentedalongwithimagingtechniques.A
chapter on the newly developed methods to culture the chordate Amphioxus provides the
transition between invertebrate and vertebrate systems. Amphibians and fishes are repre-
sented by three chapters discussing Xenopus and zebrafish embryology techniques and a
new FOP model in zebrafish. Several chapters discuss approaches using genetically engi-
neered mouse models and drugs to study BMP receptor function and activities at the
molecularandmicroscopiclevels.Thethreeterminalchaptersdiscusspreclinicalapproaches
tounderstandingBMPfunctioninboneregenerationandcancersandusingamousemodel
ofFOP.
Finally,Iwishtoexpressmygratitudetotheauthorswhocontributedgreateffortand
valuable time to this volume. Their cooperation, hard work, and the multitude of useful
v
vi Preface
experimentalhintsshouldmakethisbookahelpfulBMPresourceformanyyears.Iamtruly
indebtedtoeachcontributor.IalsomustthankDr.JohnM.Walker,firstfor theinvitation
to edit this volume and then for his patience and advice. I am particularly grateful for his
willingness to wait for “just one more exciting protocol.” May this collection promote
greater understandingoftheseessentialproteins.
Newark,NJ,USA MelissaB.Rogers,Ph.D.
Reference
1. Feng,X.-H.,Xu,P.,Lin,X.(Eds.)(2016)TGF-βsignaling: methodsin molecularbiology.Springer,
NewYork
Contents
Preface ..................................................................... v
Contributors................................................................. ix
1 CommonCellLinesUsedtoStudyBoneMorphogenetic
Proteins(BMPs)........................................................ 1
JessicaAnnCottrell,DonyaBurgess,SarahMichaels,
andMelissaB.Rogers
2 InVitroandInVivoOsteogenicDifferentiationofHuman
Adipose-DerivedStromalCells ........................................... 9
ClementD.Marshall,ElizabethA.Brett,AlessandraL.Moore,
DerrickC.Wan,andMichaelT.Longaker
3 Cell-BasedGeneTherapySystemforDeliveringBMPs ...................... 19
AustinDickerson,EleanorL.Davis,CorinneSonnet,
AlanR.Davis,andElizabethA.Olmsted-Davis
4 GenerationofEndogenousBMPTranscriptionalReporter
CellsThroughCRISPR/Cas9GenomeEditing ............................ 29
LukeD.Hutchinson,PolyxeniBozatzi,ThomasMacartney,
andGopalP.Sapkota
5 High-Throughput,Biosensor-BasedApproachtoExamineBone
MorphogeneticProtein(BMP)–ReceptorInteractions....................... 37
SenemAykulandErikMartinez-Hackert
6 MutagenesisandImagingStudiesofBMPSignaling
MechanismsinC.elegans ................................................ 51
CathySavage-Dunn,RyanJ.Gleason,JunLiu,
andRichardW.Padgett
7 GeneRegulationofBMPLigandsinDrosophila ............................ 75
BrianG.StultzandDeborahA.Hursh
8 UsingAmphioxusasaBasalChordateModeltoStudyBMP
SignalingPathway....................................................... 91
LuokWenYong,IrynaKozmikova,andJr-KaiYu
9 ProteolyticActivationofBmps:AnalysisofCleavageinXenopus
OocytesandEmbryos ................................................... 115
Hyung-SeokKim,AutumnMcKnite,andJanL.Christian
10 ImagingandQuantificationofP-Smad1/5inZebrafishBlastula
andGastrulaEmbryos................................................... 135
JosephZinski,FrancescaTuazon,YanHuang,MaryMullins,
andDavidUmulis
11 AnAdultZebrafishModelofFibrodysplasiaOssificansProgressiva............ 155
MelissaLaBontyandPamelaC.Yelick
vii
viii Contents
12 GenerationandIdentificationofGeneticallyModifiedMice
forBMPReceptors...................................................... 165
JingwenYangandYujiMishina
13 PhenotypicAnalysesofGeneticallyModifiedMiceforBMPReceptors........ 179
JingwenYangandYujiMishina
14 ImmunofluorescentVisualizationofBMPSignalingActivationon
Paraffin-EmbeddedTissueSections ....................................... 191
MaaikeAlkema,Marie-Jose´Goumans,andBoudewijnP.T.Kruithof
15 SpatialandQuantitativeDetectionofBMPActivityinMouse
EmbryonicLimbBuds .................................................. 201
MarceloRochaMarquesandJean-DenisBe´nazet
16 PharmacologicStrategiesforAssayingBMPSignalingFunction.............. 221
TeresaDinter,GeoffreyA.Bocobo,andPaulB.Yu
17 BoneMorphogeneticProteins(BMPs)andBoneRegeneration .............. 235
DeboleenaKanjilalandJessicaAnnCottrell
18 HeterotopicOssificationinMouseModelsofFibrodysplasia
OssificansProgressiva ................................................... 247
SalinA.ChakkalakalandEileenM.Shore
19 Double-HumanizedMouseModeltoStudyBone
MorphogeneticProtein(BMP)SignalinginTumorXenografts............... 257
JennaNewman,RachelNeMoyer,AndrewZloza,
andJohnLangenfeld
Index ...................................................................... 263
Contributors
MAAIKEALKEMA (cid:2) DepartmentofCellandChemicalBiology,LeidenUniversityMedical
Center,Leiden,TheNetherlands
SENEMAYKUL (cid:2) DepartmentofBiochemistryandMolecularBiology,MichiganState
University,EastLansing,MI,USA
JEAN-DENIS BE´NAZET (cid:2) DepartmentofOrofacialSciencesandPrograminCraniofacial
Biology,UniversityofCalifornia,SanFrancisco,SanFrancisco,CA,USA
GEOFFREYA.BOCOBO (cid:2) DivisionofCardiovascularMedicine,DepartmentofMedicine,
BrighamandWomen’sHospital,HarvardMedicalSchool,Boston,MA,USA
POLYXENIBOZATZI (cid:2) MRCProteinPhosphorylationandUbiquitylationUnit,SirJames
BlackCentre,UniversityofDundee,Dundee,Scotland,UK
ELIZABETHA.BRETT (cid:2) DepartmentofPlasticandHandSurgery,TechnicalUniversity
Munich,IsmaningerStrasse,Munich,Germany
DONYABURGESS (cid:2) BiologicalSciences,SetonHallUniversity,SouthOrange,NJ,USA
SALINA.CHAKKALAKAL (cid:2) DepartmentofOrthopaedicSurgery,PerelmanSchoolofMedicine
attheUniversityofPennsylvania,Philadelphia,PA,USA;CenterforResearchinFOPand
RelatedDisorders,PerelmanSchoolofMedicineattheUniversityofPennsylvania,
Philadelphia,PA,USA
JANL.CHRISTIAN (cid:2) DivisionofHematologyandHematologicMalignancies,Departmentof
Neurobiology,AnatomyandInternalMedicine,UniversityofUtahSchoolofMedicine,Salt
LakeCity,UT,USA
JESSICAANNCOTTRELL (cid:2) BiologicalSciences,SetonHallUniversity,SouthOrange,NJ,USA
ALANR.DAVIS (cid:2) Center forCellandGeneTherapy,BaylorCollegeofMedicine,Texas
Children’sHospitalandHoustonMethodistHospital,Houston,TX,USA;Sectionof
Hematology/Oncology,DepartmentofPediatrics,BaylorCollegeofMedicine,Houston,TX,
USA;DepartmentofOrthopedicSurgery,BaylorCollegeofMedicine,Houston,TX,USA
ELEANORL.DAVIS (cid:2) Center forCellandGeneTherapy,BaylorCollegeofMedicine,Texas
Children’sHospitalandHoustonMethodistHospital,Houston,TX,USA
AUSTIN DICKERSON (cid:2) Center forCellandGeneTherapy,BaylorCollegeofMedicine,Texas
Children’sHospitalandHoustonMethodistHospital,Houston,TX,USA
TERESADINTER (cid:2) DivisionofCardiovascularMedicine,DepartmentofMedicine,Brigham
andWomen’sHospital,HarvardMedicalSchool,Boston,MA,USA
RYANJ.GLEASON (cid:2) DepartmentofBiology,JohnsHopkinsUniversity,Baltimore,MD,USA
MARIE-JOSE´ GOUMANS (cid:2) DepartmentofCellandChemicalBiology,LeidenUniversity
MedicalCenter,Leiden,TheNetherlands
YANHUANG (cid:2) DepartmentofAgriculturalandBiologicalEngineering,PurdueUniversity,
WestLafayette,IN,USA
DEBORAH A.HURSH (cid:2) DivisionofCellandGeneTherapy,Center forBiologicsEvaluation
andResearch,FoodandDrugAdministration,SilverSpring,MD,USA
LUKED.HUTCHINSON (cid:2) MRCProteinPhosphorylationandUbiquitylationUnit,SirJames
BlackCentre,UniversityofDundee,Dundee,Scotland,UK
DEBOLEENAKANJILAL (cid:2) RutgersUniversity,Newark,NJ,USA
ix
x Contributors
HYUNG-SEOKKIM (cid:2) DivisionofHematologyandHematologicMalignancies,Departmentof
Neurobiology,AnatomyandInternalMedicine,UniversityofUtahSchoolofMedicine,Salt
LakeCity,UT,USA
IRYNAKOZMIKOVA (cid:2) InstituteofMolecularGeneticsoftheASCR,Prague,CzechRepublic
BOUDEWIJNP.T.KRUITHOF (cid:2) DepartmentofCellandChemicalBiology,LeidenUniversity
MedicalCenter,Leiden,TheNetherlands;DepartmentofCardiology,LeidenUniversity
MedicalCenter,Leiden,TheNetherlands;NetherlandsHeartInstitute,Utrecht,The
Netherlands
MELISSALABONTY (cid:2) PrograminCell,Molecular,andDevelopmentalBiology,SacklerSchool
ofGraduateBiomedicalSciences,TuftsUniversitySchoolofMedicine,Boston,MA,USA;
DepartmentofOrthodontics,DivisionofCraniofacialandMolecularGenetics,Tufts
UniversitySchoolofDentalMedicine,Boston,MA,USA
JOHNLANGENFELD (cid:2) DivisionofCardiothoracicSurgery,RutgersCancerInstituteofNew
Jersey,RutgersRobertWoodJohnsonMedicalSchool,Rutgers,TheStateUniversityofNew
Jersey,NewBrunswick,NJ,USA
JUNLIU (cid:2) DepartmentofMolecularBiologyandGenetics,CornellUniversity,Ithaca,NY,
USA
MICHAELT.LONGAKER (cid:2) DivisionofPlasticandReconstructiveSurgery,Departmentof
Surgery,StanfordUniversitySchoolofMedicine,Stanford,CA,USA;HageyLaboratoryfor
PediatricRegenerativeMedicine,DepartmentofSurgery,StanfordUniversitySchoolof
Medicine,Stanford,CA,USA
THOMASMACARTNEY (cid:2) MRCProteinPhosphorylationandUbiquitylationUnit,SirJames
BlackCentre,UniversityofDundee,Dundee,Scotland,UK
MARCELOROCHAMARQUES (cid:2) DepartmentofMorphology,AreaofHistologyandEmbryology,
PiracicabaDentalSchool,UniversityofCampinas,Campinas,SP,Brazil;Departmentof
OrofacialSciencesandPrograminCraniofacialBiology,UniversityofCalifornia,San
Francisco,SanFrancisco,CA,USA
CLEMENT D.MARSHALL (cid:2) DivisionofPlasticandReconstructiveSurgery,Departmentof
Surgery,StanfordUniversitySchoolofMedicine,Stanford,CA,USA
ERIK MARTINEZ-HACKERT (cid:2) DepartmentofBiochemistryandMolecularBiology,Michigan
StateUniversity,EastLansing,MI,USA
AUTUMNMCKNITE (cid:2) DivisionofHematologyandHematologicMalignancies,Departmentof
Neurobiology,AnatomyandInternalMedicine,UniversityofUtahSchoolofMedicine,Salt
LakeCity,UT,USA
SARAH MICHAELS (cid:2) BiologicalSciences,SetonHallUniversity,SouthOrange,NJ,USA
YUJIMISHINA (cid:2) DepartmentofBiologicandMaterialsSciences,SchoolofDentistry,
UniversityofMichigan,AnnArbor,MI,USA
ALESSANDRA L.MOORE (cid:2) DivisionofPlasticandReconstructiveSurgery,Departmentof
Surgery,StanfordUniversitySchoolofMedicine,Stanford,CA,USA;Departmentof
Surgery,BrighamandWomen’sHospital,Boston,MA,USA
MARYMULLINS (cid:2) DepartmentofCellandDevelopmentalBiology,UniversityofPennsylvania
PerelmanSchoolofMedicine,Philadelphia,PA,USA
RACHELNEMOYER (cid:2) DepartmentofSurgery,RutgersRobertWoodJohnsonMedicalSchool,
Rutgers,TheStateUniversityofNewJersey,NewBrunswick,NJ,USA