Nanoparticle-Based Gene Delivery Systems

for Anti-Inflammatory

Therapy

Michelle Gee, YSP Student, Wellesley High School

Yasmeen Elaywan, YSP Student, Boston Latin School

Dr. Huyen Tran, Postdoctoral Research Associate, Northeastern University

Prof. Mansoor Amiji, Department of Pharmaceutical Sciences, Northeastern

University

Inflammatory Diseases

Cancer

Pulmonary disease

Rheumatoid arthritis

Type II diabetes

Fatty liver disease

Inflammatory bowel disease

Atherosclerosis

Alzheimer’s Parkinson’s

Chronic Inflammati

on

Macrophages in Inflammation

• Macrophage: phagocytic immune cell• Can be stimulated to inflammatory M1 or anti-inflammatory M2• In inflammatory diseases macrophages are prominently M1 • Stimulate macrophages to M2 as possible treatment

M1 M2

Bosurgi et al., Front Immunol. 2011; 2: 62.

Why miRNA-223 and Nanoparticles?

• miRNA-223• Inhibits expression of pro-inflammatory cytokines• Verify that miR-223 converts macrophages to

anti-inflammatory M2 • Nanoparticles

• Protect miRNA and allow it to enter cell• Targeted delivery- HAPEI

Cy5-labeled miRNA-ve charge

HAPEI-miRNA

JJJ

Methods

I. Nanoparticle Preparation and Characterization

II. Peritoneal Macrophage Isolation & Culture

III. Peritoneal Macrophage Transfection with miRNA-223

Nanoparticles

IV. Polarization Study IV. Anti-Inflammatory Effect Study

Nanoparticle Characterization

235 d nm

Z-Average: 200.6 d nm

25 bp

Ladd

er

miR

NA

HAPEI/miR

NA

HAPEI/miR

NA/PAA

A

B

C

Uptake Study in Peritoneal Macrophages

DAPI-nuclei Cy5-HAPEI Merged

Con

trol

1 h

3 h

5 h

10 um

10 um

10 um

10 um

miRNA-223 Levels in Peritoneal Macrophages

Taqman assay

miR

-223

exp

ress

ion

(Fol

d ch

ange

com

pare

d to

unt

reat

ed)

0

50

100

150

20024h48h

Blank-HAPEI

Untreated

HAPEI/miR

NA-223

Lipofectamine/miRNA-223

Taqman Assay

Polarization Study

iNOS

Control M1 LF-miR HA-PEI/miR

Fol

d ch

ange

com

pare

d to

con

trol

0

200

400

600

800

1000

1200

Arg

Control M1 LF-miR HA-PEI/miR

Fol

d c

hang

e co

mp

are

d to

con

trol

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

iNOS(M1 inflammatory

marker)

Arg(M2 anti-inflammatory

marker)

M2M1

Untreated Macrophage

LPSIFN-γ

HAPEI/miRNA Nanoparticle

qPCR

Inflammatory Cytokine Expression Levels in

Peritoneal Macrophages

IL-1β Expression Levels IL-6 Expression Levels

TNF-α Expression LevelsLPS(inflammatory)

M2

Untreated Macrophag

e

HAPEI/miRNA Nanoparticle

qPCRM2

Conclusions and Further Research

• Conclusions• Nanoparticles prepared and entered cells• miRNA-223 treated cells expressed lower

levels of inflammatory cytokines and polarized from M1 to M2

• Promising treatment for inflammatory diseases

• Future Research• In vivo studies• Disease markers

Acknowledgements•Amiji Lab

• Professor Mansoor Amiji• Dr. Huyen Tran• Dr. George Matthaiolampakis• Swathi Krishnan• Dandan Ling• Amit Mirchandani• Yue “Jerry” Zhang

• Northeastern University Young Scholars Program

• Claire Duggan• Maureen Cabrera• Madeline Leger

• Supported by: