Shiladitya Sengupta, M.D., Assistant Professor of Medicine and Health Sciences and Technology, Harvard Medical School, Brigham & Women's Hospital: Personalized Medicine Approaches in Oncology. Presented at New Frontiers in the Management of Solid and Liquid Tumors hosted by the John Theurer Cancer Center at Hackensack University Medical Center. jtcancercenter.org/CME

1. Nanomedicine‐based
pp gy
approaches in Oncology
Dr. Shiladitya Sengupta
Harvard Medical School
Brigham and Women’s Hospital
Dana Farber Cancer Institute
Harvard‐MIT Division of Health Sciences and Technology

3. Someone diagnosed with cancer
Someone diagnosed with cancer
50 during this talk!
every second .

4. 1971

5. How does
How does
cancer
progress?

6. DNA binders that induce cell death
NH2 Cl
Pt
Cl
NH2
Cisplatin
Doxorubicin
Bleomycin

7. Tubulin inhibitors that prevent the cells from dividing
Vincristine
Taxol

8. EGFR C‐KIT C‐MET VEGFR
Molecular medicine ‐ targeting oncogenic mutations

9. USD 7000
USD 7000
per dose
USD 4500
p
per dose
USD 4300
per dose
per dose

15. earlier now
Cost of Development 250M 1.5B
Time to Market
Time to Market 10 Yrs
10 Yrs 15 Yrs
15 Yrs
NCEs ~40 ~30
Most Blockbuster Patents expire in next 2‐5 years
Most Blockbuster Patents expire in next 2 5 years

16. Cancer is more than
3 stories
3 stories
j
just a dividing cell
g

20. 2D or 3D, context matters
2D or 3D context matters
Non‐metastatic Metastatic
Weaver and Bissell

21. Endothelial cells on 2D or 3D
Endothelial cells on 2D or 3D

22. Endothelial cells Endothelial Endothelial
in MCF10A
in MCF10A cells in MCF7
cells in MCF7 cells in MDA
cells in MDA
(normal epithelium) (non‐metastatic) (highly‐metastatic)

26. P*AKT(S473)
Green = MDA‐MB‐231
Red P*AKT (S473)
Red = P AKT (S473)
Blue = DAPI

27. P*FAK Tyr925
Green = MDA‐MB‐231
Red P*FAK (Tyr925)
Red = P FAK (Tyr925)

28. OX
Uronic Acid
O
COO‐ OX
O
O O
OH NHY
O
OX Glucosamine
2 O position of uronic acid
2’O position of uronic acid
6’O and 3’O position of glucosamine
N‐position of glucosamine can either be sulfated,
N‐position of glucosamine can either be sulfated
acetylated, or unsubstituted

29. Post‐synthetic modification of HSGAGs
OH COOH OH COOH
O O N‐deacetylase/
O O
N sulfotransferase
N‐sulfotransferase
OH OH OH OH
O O O O
NAc OH NSO4 ‐2 OH
OH
O O
COOH
OH
O OH
O
O O
NSO4‐2 OH
OSO4‐2
O O
2‐O Sulfotransferase COOH
3 O Sulfotransferase
3‐O Sulfotransferase OSO4‐2 OH
6‐O Sulfotransferase O O
NSO4‐2 OSO4‐2

31. Formation of endothelial cells de novo,
Vasculogenesis
PI (Red) vWF (green)
Day 3
Day 7
D 7
Day 10

32. Characterization of HSGAG disaccharides: Sulfation increases
with endothelial differentiation
Absorbance units
Tri‐sulfate

33. Knock‐down of NDST1 disrupts neovascularization
zNDST1‐Mo
3.5 pmol

34. IGF2R GRN
GRN
MDM2
RNA Microarray Analysis
TP53
Hsp90
HSPAB MDM2
RRM2
HSPA8 GADD45A IGF2
HSP00AB1 FOXO3A
IGF2R IGF2
Hsp90 PPID RRM2B
FOXO3A CCNG1
GPI
TP73
HSP90AB1 Cav1/3 MDM2 peptidase D
GADD45 C2 zgc:76878 SESN1
TP73L
wu:fb59a01
HTRA2
zgc:110001
CAV1 TP53 selenoprotein W 2a
W,
CASP8 CLDN4 PPIA fibrinogen alpha chain BAX
6 somite stage C3
S100A8
wu:fd20g07
Dr_24890_1_S1_at
Dr_19402_1_A1_at RP527
SFRP1 PRDX1
Dr_1689_2_A1_a_at
zgc:55364
PARP2 FOS fatty acid binding pro
MMP2 nuclear receptor subfa
S100A9 FOS
cytoglobin MIF
tubulin, alpha 1
ITGAM
CTH
proteolipid protein 1a
wu:fb55b11
24 hours-po p
SFRP4 mesogenin 1 CASP8
PI3K1 FST PTPRN
wu:fc19g03
wu:fd50h12
fertilization
MBOAT5
Dr_22517_1_S1_at
PSMA2
wu:fc49d01 PARP2
FGA PTPRN2 HSPA5 matrix metalloproteina
6‐somites 20‐somites
FGG
28‐somites (24 hpf)
Dr_19794_1_A1_at
membrane protein, palm
NR2F1
growth arrest and DNA-
CLU
A2Minsulin-like growth fa
angioblasts migrate to midline
migrate to midline
MMP13 tubes assemble
tubes assemble LPA onset of angiogenesis
onset of angiogenesis
Dr_26003_1_A1_at
PSMB4
pleckstrin homology li
homology-li
CASP8 wu:fc92e10
wu:fi04f09
Dr_15033_1_S1_at
MMP2 FOS caspase 8
PARP2 20 somite stage
cyclin G1
Dr_7787_1_S1_at
poly (ADP-ribose) poly
GTP binding protein 1,
FST
SFRP1 BAX Symbol Legend murine double minute 2
forkhead box O5
zgc:92153
wu:fb96a10
tumor protein p53
TP73L wu:fc84a08
wild‐type
FOXO3A NDST1 Mo
MDM2
wild‐type
Enzyme NDST1 MoNuclear
Receptor
zgc:55750
Kinase
zgc:56722
wild‐type
wu:fj10e08
Acts on
TP73 v-fos FBJ murine osteo
wu:fj64h06
GADD45A heat shock protein 90-
Cytokine shock cognate 70-
heat
Binds
Transcription
Other
IGF2R CCNG1 Factor
IGF2 Indirect
Transmembrane
Phosphorase Peptidase Receptor Interaction
-3.00
-2.30
-1.70
-1.00
-0.33
0.33
1.00
1.70
2.30
3.00
24 hpf stage

35. Cross talk between microenvironment and endothelial
p
precursors
Foxo3A
shRNA
* control NDST1 IGF2 IGF1
Foxo3A
NDST1 IGFR1
Actin
*
control shNDST1 picropodophyllin
4 TIE2
LY294002 PI3K FOXO3
PI3K FOXO3a
3 *
AKT
Foxo3A
Rn
2
rapamycin mTOR
1
* VEGF
0
control
l
A
shFoxo3A Vasculogenesis
ro
N
HIF1
nt
R
co
sh
3A
xo
Fo

38. Story 2. The story of
Story 2 The story of
John Cossman
J h C

41. Control (30 h) Combretastatin (30 h) Doxorubicin (30 h)

42. (A) HO
O CH3 O CH3
O OH
Combretastatin (102 g)
O
O
O
n
O PLGA 5050 DL 4A
(C) Doxorubicin (g)
*
1. pNC, pyridine, CH2Cl2 8 *
leased
*
2. Doxorubicin, DMF, TEA
6 *
Total Drug Rel
O OH O
OH
4 * #
OH # #
# #
O O OH O
2 #
H3C
O O
O CH3 O CH3
H3C
NH
O
O
O
O
OH 0
OH 0 30 60 90 120 125 225 325
Doxorubicin - PLGA conjugate
O
n
O
200 Time (hours)
emulsion-solvent
nm
200
evaporation
Diameter (nm)
(B) Ultracentrifugation, sizing and
phospholipid membrane 150
coating. Combretastatin
encapsulated in lipid layer.
100
D
H3CO
50
H3CO
na le
ll
ce
c
rti
no
OH
pa
OCH3
no
na
OCH3

43. Normal blood vessels Tumor blood vessels

44. (A)
TUNEL/HIF1a vWF
Vehicle L[C] Vehicle L[C]
Ve
ehicle
NC[D] L[CD] L[CD]
NC[D]
L[
[C]
NC[D]+L[C] NC
NC[D]+L[C] NC
NC (ld)
NC
C[D]
Lewis lung carcinoma B16/F10 melanoma
NC[
5500 Lewis lung carcinoma
10000 B16/F10 melanoma
[D]+L[C]
5000
7500
4500 5000
3
3
olume mm
olume mm
4000 2500
3500
2000
3000
Tumor Vo
Tumor Vo
NC
2500 1500
2000
1500 1000
1000
500
500
L[CD]
0 0
9 11 13 15 17 9 11 13 15 17
Days Days
Veh L[C] NC[D] L[CD]
NC[D]+L[C] NC NC( ld)

45. Coming back to John
Coming back to John

46. Some results from Phase 1, currently Phase 2

47. Story 3. The future: design of
y g
nanomedicine based on SAR

48. Understanding SAR to design a nanomedicine
Cl OH
Cl Cl
aquation
q DNA
binding
Cisplatin
Carboplatin Oxaliplatin

50. Understanding SAR to design a nanomedicine
DMF, H2O
Dry DMF,
DBU
umber %
30
pH<7
20
Nu
10
0
1 10 100 1000 10000 pH>7 OH
Size (d.nm) n n OH OH
HO
H
O O O N O
HO
O HN O O
OH
Pt OH Pt
H 3N NH3 OH
O H 3N NH3
‐1611.54
‐2210
‐
‐500 ‐1500 ‐2500
ppm

51. Lung Cancer
g
PIMA‐GA‐ PIMA‐GA‐
Ovarian Cancer Cisplatin
(3mg/kg)
CisPt CisPt Cisplatin
(1.25 mg/kg)
(3mg/kg) (1.25 mg/kg) Vehicle
PIMA GA CisPt
PIMA‐GA‐CisPt [1.25mg/kg] PIMA‐GA‐CisPt [1.25mg/kg]
PIMA‐GA‐CisPt [3 mg/kg] PIMA‐GA‐CisPt [3 mg/kg]
(C) CisPt [1.25 mg/kg] CisPt [1.25 mg/kg]
CisPt [3mg/kg] CisPt [3mg/kg]
Vehicle Vehicle
PIMA-GA-CisPt PIMA-GA-CisPt
Vehicle Control CisPt
Ci Pt [3 mg/kg]
/k ] 225
[1.25 mg/kg] [3 mg/kg] 200
175
Kidney
150
mg
125
***
100
75
50
25
Tumor
0
)
)
5)
)
e
kg
kg
(3
cl
.2
is
hi
g/
g/
(1
C
ve
5m
m
is
A-
(3
C
.2
G
A-
is
(1
A-
C
G
M
is
A-
PI
C
M
PI

52. Translating it to the clinics
(a) (b)
(a) Cisplatin Cisplatin‐NP
Control (3mg/kg) (3mg/kg)
tment
(a)
( )
LLC 4T1 CP20
Pretreat
(b) (c)
ent
Post treatme
(b)
‐1621.5
(f)
10 100 1000

53. (c) Cisplatin (1 mg/kg) Cisplatin (3 mg/kg) Cisplatin (1 mg/kg) Cisplatin (3 mg/kg)
Cisplatin NP (1 mg/kg) Cisplatin NP (3 mg/kg) Cisplatin NP (1 mg/kg) Cisplatin NP (3 mg/kg)

54. Metastasis
What drives cancer?
SO4

55. Abhimanyu Paraskar Funding Support
DoD Postdoctoral Scholar
Cisplatin‐nanoparticle
l l NIH
DoD
Coulter Foundation
Sudipta basu American Heart Association
Charles A King Postdoctoral Scholar
Charles A King Postdoctoral Scholar Mary Kay Ash Foundation
y y
Nano‐biotechnology CIHR
Charles A King Trust
Rania Harfouche
f h
CIHR Postdoctoral Scholar
Vasculogenesis
Yamicia Connor
NIH‐ MD,PhD Scholar
Tumor Microenvironment
Stephanie Piecewicz
Harvard‐MIT PhD
Harvard MIT PhD
Vasculogenesis

56. Shape affects outcome