Pharmacogenetics of microRNAs and microRNAs biogenesis machinery in pediatric Acute Lymphoblastic Leukemia Elixabet López-López, Ángela Gutierrez-Camino,

Pharmacogenetics of microRNAs and microRNAs biogenesis machinery in

pediatric Acute Lymphoblastic Leukemia

Elixabet López-López, Ángela Gutierrez-Camino, Idoia Martín-Guerrero, M Ángeles Piñán, Nagore García de Andoin, Aurora Navajas, Purificación

García-Miguel, Javier Ballesteros, África García-Orad

1

Acute Lymphoblastic Leukemia

Most common pediatric malignancy

Disorder of the lymphoblastsAccumulation of blast cells in BM and PB

IntroductionIntroduction

Normal

ALL

2

Survival 10% 80%

Risk groups characterization

Well-established treatment protocols directed to risk groups


3

DNR VCR

PDN CFM

ASP TIT (MTX)

MTX 6MP

AraC TIT (MTX)

VCR PDN ASP

MTX 6MP

CFM TIT (MTX)

epiADR VCR

DEXA CFM

MTX ASP

AraC TIT (MTX)

VCR PDN

MTX 6MP

TIT (MTX)

VCR DNR

DEXA CFM

MTX ASP

AraC TIT (MTX)

Induction Consolidation

Intensification

Maintenance

HR

VHR

Induction

Induction

Consolidation

Consolidation Blocks Intensification

Maintenance

Graft

MTX 6MP

AraC TIT (MTX)

MTX 6MP

AraC TIT (MTX)

DNR VCR

PDN CFM

ASP TIT (MTX)

DNR VCR

PDN CFM

ASP TIT (MTX)


SR

4

LAL/SHOP protocol

DNR VCR

PDN CFM

ASP TIT (MTX)

MTX 6MP

AraC TIT (MTX)

VCR PDN ASP

MTX 6MP

CFM TIT (MTX)

epiADR VCR

DEXA CFM

MTX ASP

AraC TIT (MTX)

VCR PDN

MTX 6MP

TIT (MTX)

VCR DNR

DEXA CFM

MTX ASP

AraC TIT (MTX)


Intensification

Maintenance

HR

VHR

Induction

Induction

Consolidation


Maintenance

Graft

MTX 6MP

AraC TIT (MTX)

MTX 6MP

AraC TIT (MTX)

DNR VCR

PDN CFM

ASP TIT (MTX)

DNR VCR

PDN CFM

ASP TIT (MTX)


SR

5

LAL/SHOP protocol

DNR VCR

PDN CFM

ASP TIT (MTX)

MTX 6MP

AraC TIT (MTX)

VCR PDN ASP

MTX 6MP

CFM TIT (MTX)

epiADR VCR

DEXA CFM

MTX ASP

AraC TIT (MTX)

VCR PDN

MTX 6MP

TIT (MTX)

VCR DNR

DEXA CFM

MTX ASP

AraC TIT (MTX)


Intensification

Maintenance

HR

VHR

Induction

Induction

Consolidation


Maintenance

Graft

MTX 6MP

AraC TIT (MTX)

MTX 6MP

AraC TIT (MTX)

DNR VCR

PDN CFM

ASP TIT (MTX)

DNR VCR

PDN CFM

ASP TIT (MTX)


SR

6

Toxicity- Requires treatment reduction

LAL/SHOP protocol

DNR VCR

PDN CFM

ASP TIT (MTX)

MTX 6MP

AraC TIT (MTX)

VCR PDN ASP

MTX 6MP

CFM TIT (MTX)

epiADR VCR

DEXA CFM

MTX ASP

AraC TIT (MTX)

VCR PDN

MTX 6MP

TIT (MTX)

VCR DNR

DEXA CFM

MTX ASP

AraC TIT (MTX)


Intensification

Maintenance

HR

VHR

Induction

Induction

Consolidation


Maintenance

Graft

MTX 6MP

AraC TIT (MTX)

MTX 6MP

AraC TIT (MTX)

DNR VCR

PDN CFM

ASP TIT (MTX)

DNR VCR

PDN CFM

ASP TIT (MTX)


SR

7

Toxicity- Requires treatment reduction

PREDICTIVE MARKERS

LAL/SHOP protocol


ABCB1

ABCG2ABCC2

MTX MTXSLCO1A2

ABCC3ABCC2

ABCC4

SLC22A6 SLC22A8

SLC19A1

SLCO1B3

ABCC1ABCG2

LIVER CELL

KIDNEY CELL

Elimination

Elimination

Via bileVia bloodstream

SLCO1B1

8

384 SNPs151 B-ALLLAL/SHOP

Gene SNP Allele OR (CI 95%) p Function

ABCC4 rs9516519 C 0.17 (0.05-0.5) 0.0003086(0.0188)

Putative miRNA binding site

ABCC2 rs3740065 C 3.75 (1.6-8.79) 0.002009(0.0340) Putative intronic enhancer

9

E. Lopez-Lopez et al. Pharmacogenetics and Genomics 2013



ABCC4 rs9516519 C 0.17 (0.05-0.5) 0.0003086(0.0188)



10




ABCC4 rs9516519 C 0.17 (0.05-0.5) 0.0003086(0.0188)



11




ABCC4 rs9516519 C 0.17 (0.05-0.5) 0.0003086(0.0188)



12



ABCB1

ABCG2

SLCO1A2

ABCC3

SLC22A6 SLC22A8

SLC19A1

SLCO1B3

ABCC1ABCG2

LIVER CELL

KIDNEY CELL

Via bile

Via bloodstream

SLCO1B1

ABCC2

MTX MTX

ABCC2

ABCC4Elimination

Elimination

MicroRNAs

1-BINDING

13


MicroRNAs


1-BINDING

2-LEVEL

14

MicroRNAs


1-BINDING

2-LEVEL

15Adapted from Ryan et al, 2010

MicroRNAs


1-BINDING

2-LEVEL

16Adapted from Ryan et al, 2010

To detect new toxicity markers in pediatric B-ALL patients studying miRNA-related polymorphisms

AimAim

Material and methodsMaterial and methods

1. miRNAs biogenesis and processing genes: 1. MAF: 5% 2. Functional SNPs 3. bibliographic

2. premiRNAs: 1. MAF: 1%

118 SNPs

18

152 children with B-ALLLAL/SHOP

Taqman Openarray

21 Genes 74 SNPs

19

Results and DiscussionResults and Discussion

Gene SNP Toxicity Phase Genotype No toxicity n (%)

Toxicity n (%)

OR (95% CI) p-value

DROSHA rs639174 Vomits Cons CC 47 (92.2) 4 (7.8) 1.00 0.0003*(0.031)CT/TT 43 (65.2) 23 (34.9) 6.28 (2.01-19.64)

rs2287584 Vomits Cons TT 54 (88.5) 7 (11.5) 1.00 0.0026CT/CC 41 (66.1) 21 (33.9) 3.95 (1.53-10.18)

Vomits Ind TT 41 (65.1) 22 (34.9) 1.00 0.0041CT/CC 54 (80.6) 13 (19.4) 0.45 (0.20-1.00)

rs10035440 Hyperbilirubinemia Ind TT 43 (72.9) 16 (27.1) 1.00 0.0088CT/CC 65 (92.9) 5 (7.1) 0.23 (0.08-0.68)

Diarrhea Cons TT/CT 104 (94.6) 6 (5.5) 1.00 0.0091CC 9 (75.0) 3 (25.0) 5.78 (1.23-27.06)

rs4867329 Vomits Cons AA/AC 74 (72.6) 28 (27.5) 1.00 0.0137CC 21 (95.5) 1 (4.6) 0.13 (0.02-0.98)

rs3805500 Hepatic toxicity Cons AA/AG 70 (68.0) 33 (32.0) 1.00 0.0259GG 17 (94.4) 1 (5.6) 0.12 (0.02-0.98)

Vomits Cons AA 41 (89.1) 5 (10.9) 1.00 0.0279AG/GG 53 (70.7) 22 (29.3) 3.40 (1.19-9.76)

rs10719 Vomits Cons GG 56 (83.6) 11 (16.4) 1.00 0.0266AG/AA 35 (66.0) 18 (34.0) 2.62 (1.11-6.19)

MTX plasma levels Cons GG/AG 77 (63.6) 44 (36.4) 1.00 0.0359AA 9 (100) 0 (0.0) NE (NE-NE)

rs7735863 Hepatic toxicity Cons GG 58 (66.7) 29 (33.3) 1.00 0.0405AG/AA 30 (85.7) 5 (14.3) 0.33 (0.12-0.95)

rs6877842 Diarrhea Ind GG 73 (92.4) 6 (7.6) 1.00 0.0411GC/CC 39 (79.6) 10 (20.4) 3.12 (1.05-9.23)

Hepatic toxicity Cons GG 59 (77.6) 17 (22.4) 1.00 0.0455CG/CC 27 (60.0) 18 (40.0) 2.31 (1.04-5.17)




Rotunno et al, 2010


Rotunno et al, 2010


Rotunno et al, 2010


Gene SNP Toxicity Genotype Absence n (%) Presence n (%) OR (95% CI) p

mir-453 rs56103835 VomitsAA 66 (83.5) 13 (16.5) 1.00

0.0141GA/GG 28 (63.6) 16 (36.4) 2.90 (1.23-6.82)

MTX clearance

AA 62 (72.1) 24 (27.9) 1.000.0297

GA/GG 25 (53.2) 22 (46.8) 2.27 (1.08-4.77)

mir-2053 rs10505168 MucositisTT 43 (82.7) 9 (17.3) 1.00

0.0154CT/CC 68 (95.8) 3 (4.2) 0.21 (0.05-0.82)

mir-1206 rs2114358 Mucositis

AA/AG 96 (92.3) 8 (7.7) 1.00

0.0254GG 15 (75.0) 5 (25.0) 4.57 (1.28-16.28)

Diarrhea

AA/AG 98 (95.2) 5 (4.9) 1.00

0.0365GG 16 (80.0) 4 (20.0) 4.90 (1.49-20.21)

mir-604 rs2368393Renal toxicity

AA 65 (95.6) 3 (4.4) 1.00

0.0271AG/GG 47 (83.9) 9 (16.1) 4.15 (1.07-16.15)

mir-1294 rs13186787Hyperbilirubinemia

AA 110 (93.2) 8 (6.8) 1.00

0.0424AG 3 (60.0) 2 (40.0) 9.17 (1.33-63.01)

mir-2110 rs17091403 VomitsCC 84 (80.0) 21 (20.0) 1.00

0.0471CT 11 (57.9) 8 (42.1) 2.91 (1.04-8.14)

42 pre-miRNAs 44 SNPs

25

Gene SNP Toxicity Genotype Absence n (%) Presence n (%) OR (95% CI) p

mir-453 rs56103835 VomitsAA 66 (83.5) 13 (16.5) 1.00

0.0141GA/GG 28 (63.6) 16 (36.4) 2.90 (1.23-6.82)

MTX clearance

AA 62 (72.1) 24 (27.9) 1.000.0297

GA/GG 25 (53.2) 22 (46.8) 2.27 (1.08-4.77)

mir-2053 rs10505168 MucositisTT 43 (82.7) 9 (17.3) 1.00

0.0154CT/CC 68 (95.8) 3 (4.2) 0.21 (0.05-0.82)

mir-1206 rs2114358 Mucositis

AA/AG 96 (92.3) 8 (7.7) 1.00

0.0254GG 15 (75.0) 5 (25.0) 4.57 (1.28-16.28)

Diarrhea

AA/AG 98 (95.2) 5 (4.9) 1.00

0.0365GG 16 (80.0) 4 (20.0) 4.90 (1.49-20.21)

mir-604 rs2368393Renal toxicity

AA 65 (95.6) 3 (4.4) 1.00

0.0271AG/GG 47 (83.9) 9 (16.1) 4.15 (1.07-16.15)

mir-1294 rs13186787Hyperbilirubinemia

AA 110 (93.2) 8 (6.8) 1.00

0.0424AG 3 (60.0) 2 (40.0) 9.17 (1.33-63.01)

mir-2110 rs17091403 VomitsCC 84 (80.0) 21 (20.0) 1.00

0.0471CT 11 (57.9) 8 (42.1) 2.91 (1.04-8.14)


42 pre-miRNAs 44 SNPs

TargetsABCC1, ABCC2, ABCC4

26

miRNA-related SNPs could be useful toxicity markers

ConclusionConclusion

Conclusión

27

A. García-OradI. Martín GuerreroA. Gutiérrez-CaminoN. BilbaoM. PombarZ. Askaiturrieta

J. BallesterosB. Santos

A. NavajasMA. Piñán

P. García-Miguel

J. UrizN. García de Andoin

AcknowledgementsAcknowledgements

Documents

Pharmacogenetics of microRNAs and microRNAs biogenesis machinery in pediatric Acute Lymphoblastic Leukemia Elixabet López-López, Ángela Gutierrez-Camino,