Investigate Cellular Responses for Investigate Cellular Responses for Environmental CuesEnvironmental Cues

Qian Wang

Department of Chemistry and Biochemistry, and NanocenterUniversity of South Carolina, SC, USA

Cell shape determines fate of stem cellsCell shape determines fate of stem cells

Mrksich M., et al. PNAS, 2010, 16, 4872-4877

Star shape and flower shape expressed different differentiation profile

Flower shape promoted adipogenesisStar shape promoted osteogenesis

Flower

Star

ECM Mimetic SubstratesECM Mimetic Substrates

Abrams, G. A., et al., Cell Tissue Res , 2000, 299, 39-46 Kim, D. H. et al., PNAS , 2010, 107,565-570Roohani-Esfahani, S.I., et al., Biomaterials , 2010, 31, 5498-5509 Pham, Q. P. et al., Tissue Eng , 2006, 12, 1197-211

50um

Natural ECM architecture

ECM mimetic substrates

Nature 414:105, 2001

Models of Tumor HeterogeneityModels of Tumor Heterogeneity

Are We Targeting the Right Cancer Cells?Are We Targeting the Right Cancer Cells?

Sarkar, B. ; Dosch, J., et al. Chem. Rev. 2009, 109, 3200–3208

Major Challenges:•Lack of universal biomarker•Lack of In vitro cell culture model

Major Challenges:•Lack of universal biomarker•Lack of In vitro cell culture model

Nature 414:105, 2001

Dr. Hexin ChenBiologist

Dr. Xinfeng LiuMathematician

A Novel Breast Cancer Stem Cell Model - A Novel Breast Cancer Stem Cell Model - H6O5 CellsH6O5 Cells

Lo, P. K., et al., Oncogene 2012, (doi: 10.1038/onc.2011.439)

Her2/neu induced mouse mammary tumor

Consists of a highly tumorigenic population (CSCs)

Isolated based on two integrin markers CD49f and CD61

Confirmed by its in vivo tumorigenicity

NMuMG H6O5

CD61

CD

49

f G1G2

G3

G1: CD49fhighCD61high

G2: CD49fhighCD61low

G3: CD49flowCD61low

1.2 89.9

6.2 2.7

CD49fhighCD61high

2.7 74.0

8.0 15.2

CD49fhighCD61low

2.0 25.4

35.3 37.3

CD49flowCD61low

CD24

ES

A

CD

49f

CD61

Fluorescence assisted cell sorting

H6O5

Aligned fiber substrate

Tissue culture well

H6O5 Cells on Fiber SubstrateH6O5 Cells on Fiber Substrate

• Morphology

• EMT- associated gene expression

Culture for 3- 5 days

Morphology of Electrospun Fiber Morphology of Electrospun Fiber SubstrateSubstrate

SEM images of aligned (d = 1.8 ± 0.46 μm) and random (d = 2.0 ± 0.65 μm) Scale bar = 20 μm

Poly-Caprolactone

BiodegradableCytocompatible

AAligned Fiber Random Fiber

Saha. S et al.,Langmuir, 2012, 28, 2028-2034

Morphology of H6O5 Cells on Aligned Morphology of H6O5 Cells on Aligned Fiber SubstrateFiber Substrate

10x

10x

• Cell attach to fiber• Elongated morphology

• Growth, • Proliferation • Maintain elongated morphology

10x

Green: actin cytoskeletonBlue: nucleiWhite arrows: underlying fiber direction,Scale bar = 40 µm

a b

S. Saha et al. Langmuir, 2012, 28, 2028-2034

Enrichment of Cancer Stem Cells Using Enrichment of Cancer Stem Cells Using Nano-fibrous ScaffoldsNano-fibrous Scaffolds

S. Saha et al. Langmuir, 2012, 28, 2028-2034

Gene expression by qRT-PCR

S. Saha et al. Langmuir, 2012, 28, 2028-2034

Enhanced Expression of EMT Markers of Enhanced Expression of EMT Markers of MCF-7 CellsMCF-7 Cells

ALDH (Aldehyde DALDH (Aldehyde Dehydrogenaseehydrogenase) Activity ) Activity of MCF-7 Cellsof MCF-7 Cells

AAligned Fiber Random Fiber

Sheng Feng

Increased Mammosphere-Forming Increased Mammosphere-Forming CapabilityCapability

AAligned Fiber Random Fiber

Sheng Feng

Increased Invasion Property of Epithelial Increased Invasion Property of Epithelial Breast Cancers Cells Cultured On PCL FiberBreast Cancers Cells Cultured On PCL Fiber

AAligned Fiber Random Fiber

Sheng Feng

Results of MDA-MB-231 Cells Cultured Results of MDA-MB-231 Cells Cultured from PCL Fiberfrom PCL Fiber

AAligned Fiber Random Fiber

Sheng Feng

A Hypothesis of the Double-Feedback to A Hypothesis of the Double-Feedback to Control the CSCs Fate Control the CSCs Fate

A

Dr. Xinfeng Liu, Hexin Chen

Zhongwei, Jianhua, Mike Bruckman O. D. Velev et al, Small 2006, 2, 1462

Alignment of Rod Like NanoparticlesAlignment of Rod Like Nanoparticles

2D Alignment of Colloidal Particles2D Alignment of Colloidal Particles

Dimitrov and Nagayama, Langmuir 1996, 12, 1303

2D Alignment of Protein Nanoparticles2D Alignment of Protein Nanoparticles

Mike Bruckman, Zhongwei Niu O. D. Velev et al, Small 2006, 2, 1462

Jianhua Rong, Zhongwei Niu, in collaboration with Dr. Charlene Mello

Controlled Alignment of Cell GrowthControlled Alignment of Cell Growth

Rosamond W et al., Circulation 2008, 117 (4): e25-146Rosamond W et al., Circulation 2008, 117 (4): e25-146

United States, 2004. Source: NCHS and NHLBI

Cardiovascular Disease in USACardiovascular Disease in USA

Vascular Disorders and Vascular Disorders and Pathogenesis of Atherosclerosis Pathogenesis of Atherosclerosis

Osterud & Bjorklid; Physiol Rev. 2003, 83, 1069-112

VSMC Proliferative formVSMC Proliferative formVSMC Normal formVSMC Normal form

Smooth Muscle Cells: Different MorphologiesSmooth Muscle Cells: Different Morphologies

Lilly

Control

AlignedScaffold

Significance of the Alignment of Significance of the Alignment of Smooth Muscle CellsSmooth Muscle Cells

Are There a Good Model to Predict/Explain Are There a Good Model to Predict/Explain The Cellular Responses to Contact Cues??The Cellular Responses to Contact Cues??

Are There a Good Model to Predict/Explain Are There a Good Model to Predict/Explain The Cellular Responses to Contact Cues??The Cellular Responses to Contact Cues??

AcknowledgmentsAcknowledgments

BNP Assembly:Tom Russell (UMass)Yao Lin (UConn)Alex Böker (MPI)P. Thiyagarajan (DOE)Byeongdu Lee (ANL)Lin Yang (BNL)Z. Su (CIAC, CAS)C. Xiaodong Li (USC)Wenke Zhang (Jilin U)

Cell StudyHexin Chen (USC)Tara Soto-Atwood (UF)J. Potts (USC)

Protein Binding StudyLukasz Lebioda (USC)Frank Berger (USC)

Virus:Theo Dreher (OSU)Rick Nelson (Noble)Charlene Mello (ARL)Peter E. Prevelige (UAB)

Polymeric Assembly:Mihaela Stephan (UTD)Yuan Lin (CIAC, CAS)

$$$NSF-CAREER/NSF-DMRNIH-COBRESloan/Camille DreyfusDOD-ARODOE-BESW. M. Keck FoundationSC EPSCoR Office & SCCC

Wang Group:L. Andrew Lee (Postdoc)Xinrui Duan (Postdoc)Honglin Liu (Postdoc)Jun Hu (Postdoc)Pan He (Research Ass.)

Dan MenascoNikki Sitasuwan Matsepo RamaboliAmie J. Luckanagul

Wang Group Alumni:Yi Chen (Res. Professor)Gary Horvath (S. Scientist)

Postdoc:K. SivakumarS. ChittaboinaV.S. KotakadiZhongwei NiuQingbing ZengCéline le DroumaguetJianhua RongYuan LinJiangli FanXingjie Zan

Undergraduate Student:Lisa AlexanderGiang Nguyen……

Sheng FengHong GuanClorissa WashingtonYixing ChengEnoch AdoglaIin LvGift MetavarayuthPeiyi WangFang Zhou (CIAC)Xia Zhao (CIAC)Limin Chen (CIAC)

Many talent undergrad.

Graduate Student:Hannah BarnhillFang XieSu LongBrandon CashSaswat KabisatpathySiqi LiMike BruckmanGagandeep KaurTao LiKai LiChao WangJerry OxsherLaying WuEve Suthiwangcharoen Sharmi SahaLilly Balizan

SimulationQi Wang (USC)Xinfeng Liu (USC)