Alteration of lipids and the transcription of lipid-related genes in Imatinib-treated CML

Martin Ellis, MD1,2, Eilon Krashin, MD3, Sarah Gan1,2, Orly Hamburger, MD1 and Osnat Ashur-Fabian, Ph.D1,2

1Translational Hemato-Oncology Laboratory, The Hematology Institute and Blood Bank, Meir Medical Center, Kfar-Saba, Israel

2Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel 3Department of internal medicine A, Meir Medical Center, Kfar-Saba, Israel

• Clonal myeloproliferative neoplasm

• Philadelphia chromosome

• BCR/ABL1 tyrosine kinase

CML Chronic Myeloid Leukemia

Imatinib

Imatinib Metabolic effects

NEJM Study pitfalls

• Small study group (n=9)

• Heterogeneous cohort

• Limited lipid-profile

• No statistical analysis

• No in-vitro study

To confirm Imatinib beneficial lipid effect

in a larger-scale group of CML patients

Study aim

Study Cohort

• 40 CML patients

• January 2005 - March 2015

• 3m of imatinib treatment

Baseline 3-12 months >12 months

Retrospective Follow-up

Before Imatinib

After Imatinib

RESULTS

• 55% male

• Mean age 67.3

• Median time from diagnosis to imatinib –1m

• 47.5% prior treatment with statins

Patient data

3-12 months:

Imatinib improves lipid profile Baseline:

>12 months:

Imatinib improves lipid profile Baseline:

Imatinib improves lipid profile All patients

Imatinib improves lipid profile No statins

Lipid profile without statins, n=21

Imatinib improves lipid profile With statins

Lipid profile with statins, n=19

To study in vitro the mechanism of

Imatinib-lipid-effect in CML

Study aim

K-562 Imatinib 24h 48h 72h 96h

CML model In vitro

Lipid-related genes transcription

CETP

Apobec1

Apobec1

LDL Receptor

LDL-R

HMGcoAR

RESULTS

Imatinib Induces CETP mRNA

0

20

40

60

80

100

120

24 48 72 96

CET

P m

RN

A e

xpre

ssio

n

(Fo

ld o

f co

ntr

ol)

Hours after 1mM Imatinib

**

*

*

*

0

2

4

6

8

10

24 48 72

Ap

ob

ec1

mR

NA

exp

ress

ion

(F

old

of

con

tro

l)

Hours after 1mM Imatinib

* **

Imatinib Induces apobec-1 mRNA

0

2

4

6

8

10

12

14

16

24 48 72 96

LDL-

R m

RN

A e

xpre

ssio

n

(Fo

ld o

f co

ntr

ol)

Hours after 1mM Imatinib

*

**

*

Imatinib Induces LDL-R mRNA

0

2

4

6

8

10

12

14

24 48 72 96 HM

Gco

AR

mR

NA

exp

ress

ion

(F

old

of

con

tro

l)

Hours after 1mM Imatinib

**

**

*

Imatinib Induces HMGcoAR mRNA

How are lipid-genes regulated by imatinib?

53p regulated genes

Novel p53 regulated genes

Apobec-1

Apo B

Novel p53 regulated genes

Genomic

location

% response

element score

Gene

name

Promoter 92.47% Apobec1

Intron 2 91.2% Apo B

Promoter 86.29% LDL

receptor

Promoter 83% HMGCoA

R

Intron 1 83% CETP

Promoter 89.1% LRP1

Promoter 83.56% Apo E

Promoter 85.38 Apo A1

Promoter 83% Apo C-I

(VLDL)

Promoter 83.4% LCAT

Promoter 88% Apo B48

receptor

Promoter 86%-Two sites Apo A2

Promoter 81-85%-Two

sites

Apo A4

Promoter 80-81%-Two

sites

Apo C2

Intron 1 86% Apo C3

Intron 1 92.5% Apo D

Promoter+intron

4

88%- 90%-

Two sites

LPL

Genomic

location

% response

element score

Gene

name

Promoter 92.47% Apobec1

Intron 2 91.2% Apo B

Promoter 86.29% LDL

receptor

Promoter 83% HMGCoA

R

Intron 1 83% CETP

Promoter 89.1% LRP1

Promoter 83.56% Apo E

Promoter 85.38 Apo A1

Promoter 83% Apo C-I

(VLDL)

Promoter 83.4% LCAT

Promoter 88% Apo B48

receptor

Promoter 86%-Two sites Apo A2

Promoter 81-85%-Two

sites

Apo A4

Promoter 80-81%-Two

sites

Apo C2

Intron 1 86% Apo C3

Intron 1 92.5% Apo D

Promoter+intron

4

88%- 90%-

Two sites

LPL

Novel p53 regulated genes

HMGcoAR LDL-R

Cholesterol

Is the lipid-genes expression in K-562

p53-mediated?

p53 mutation in K-562

mut. Primers

0

2

4

6

8

10

12

14

16

24 48 72 96

p5

3 m

RN

A e

xpre

ssio

n

(Fo

ld o

f co

ntr

ol)

Hours after 1mM Imatinib

**

**

** **

p53 is induced by imatinib

K562 (blast crisis) no p53 expression

p63/73 expression?

“lipid clearance genes” expression

Imatinib

CML In vitro

Chronic phase CML

Wild type p53

p53 expression

“lipid clearance genes” expression

Favorable lipid profile

Imatinib

CML Patients

Summary

• A favorable lipid profile with Imatinib in CML patients

• Imatinib induces lipid-clearing-genes in vitro

Future plans CML patients

• To prove the imatinib-lipid molecular association in cells collected before/after Imatinib

• To asses p53 regulation

• To assess new generation TKI’s effect

Translational Hemato-Oncology

The Hematology Institute and blood bank

• Dr. Martin Ellis, Director • Sarah Gan, MD student

• Orly Hamburger, Senior Hematologist

Thanks

• Dr. Osnat Ashur-Fabian