Paolo Ghia - Ematologialasapienza.it 19 Sett/P. Ghia.pdf · sf3b1 mutations induce common and lineage specific aberrant messenger rna splicing in malignancies including chronic lymphocytic

Chronic Lymphocytic Leukemia

Paolo Ghia

CLL – Biology

CLL - Clinical

5 + 40 (1 oral + 2 poster sessions)

5 + 21 (1 oral + 1 poster session)

Number of abstractsTopics

Lunch Debate CLL: are novel prognostic

biomarkers ready for prime time?

CLL: are novel prognostic biomarkers ready for prime time?

NO! Yes!

Gianluca Gaidano Kostas Stamatopoulos

Università Piemonte Orientale Institute of Applied BiosciencesA. Avogadro CERTH, Thessaloniki

NOTCH(NOTCH1)

Splicing(SF3B1)

NF-κB(BIRC3)

Wang et al, NEJM 2011; Quesada, Nature Genetics

2011; Puente qt, Nature, 2011; Fabbri et el, JEM 2011

TP53 M

17p-

TP53 M /17p-

Wt

TP53 abnormalities

30/318=9.4%Wt (n=277; nr)

TP53 M (n=14; 30.2 mo)

17p- (n=16; 19.2 mo)

TP53 M

17p-

TP53 M /17p-

Wt

TP53 abnormalities

46/520=8.8%

GCLLSG CLL4 UK LRF CLL4

Months

OS OS

Wt (n=474)

TP53 M (n=13)17p- (n=8)

TP53 M/17p- (n=25)

5’ 3’

1 DNA BINDING

EX4 EX9

393

Missense Nonsense Frameshift

TP53

Years

TP53 mutations are an independent and robust

predictor of treatment failure in clinical trials

Happy with p53 aberrations only?

Yes

A. True predictor and not only prognosticator

B. Availability of “overcoming” treatment options

C. More frequent than initially thought

Not reallyA. Infrequent at diagnosis – identifies a particularly aggressive MINORITY

B. DOES NOT explain more than 50% of refractory CLL

C. Things are advancing after all!

FCR

FC

Therapy:

NOTCH1:

wild type

mutated

Stilgenbauer et al. ASH 2012, Abstract 433

CLL: MRD as a Surrogate Endpoint for Clinical Trials

White Oak – February 27, 2013

Dreger P et al. Blood 2013 Schnaiter A et al. Blood 2013

SIMULTANEOUS SESSION

CLL an related disorders: Cinical II

DISEASE PROGRESSION ON IBRUTINIB THERAPY IS UNCOMMON AND IS

ASSOCIATED WITH THE ACQUISITION OF RESISTANCE MUTATIONS: A SINGLE

CENTER EXPERIENCE OF 267 PATIENTS (Maddocks, Columbus)

EFFICACY OF IDELALISIB IN CLL SUBPOPULATIONS HARBORING DEL(17P) AND

OTHER ADVERSE PROGNOSTIC FACTORS: RESULTS FROM A PHASE 3,

RANDOMIZED, DOUBLE-BLIND, PLACEBO-CONTROLLED TRIAL (Stilgenbauer,

Ulm)

IBRUTINIB INTERFERES WITH THE CELL-MEDIATED ANTI-TUMOUR ACTIVITIES

OF THERAPEUTIC CD20 ANTIBODIES: IMPLICATIONS FOR COMBINATION

THERAPY (Da Roit, Bergamo)

PHASE 2 STUDY OF THE BTK INHIBITOR IBRUTINIB IN GENETIC RISK-STRATIFIED

RELAPSED AND REFRACTORY PATIENTS WITH CHRONIC LYMPHOCYTIC

LEUKEMIA (CLL)/SMALL LYMPHOCYTIC LYMPHOMA (SLL) (Maddocks, Columbus)

SF3B1 MUTATIONS AND OUTCOME IN CLL PATIENTS TREATED WITH

CHLORAMBUCIL (CHL) OR OFATUMUMAB-CHL (O+CHL): RESULTS FROM THE

PHASE III STUDY COMPLEMENT 1 (Tausch, Ulm)

OMB110911CHL vs CHL + OFA

SF3B1mut was associated with high WBC, male sex, absence of NOTCH1mut and showed a trend to

absence of +12q and 17p-

SF3B1mut significantly associated with shorter PFS in univariate analysis but not in multivariable analysis


CLL and related disorders – Biological

ACQUIRED INITIATING MUTATIONS IN EARLY HEMATOPOIETIC CELLS OF

CHRONIC LYMPHOCYTIC LEUKEMIA PATIENTS (Damm, Villejuif)

SF3B1 MUTATIONS INDUCE COMMON AND LINEAGE SPECIFIC ABERRANT

MESSENGER RNA SPLICING IN MALIGNANCIES INCLUDING CHRONIC

LYMPHOCYTIC LEUKEMIA AND CONFER SENSITIVITY TO SPLICEOSOME

INHIBITION (Buonamici, Cambridge, US)

IL23 RECEPTOR (IL23R) IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL):

MODULATION, MICRORNA (MIRNA) REGULATION AND PREDICTION POWER. A

PROSPECTIVE MULTICENTER O-CLL STUDY (Morabito, Cosenza)

ISOTYPE MATTERS: FUNCTIONAL DIFFERENCES AFTER SIGM VERSUS SIGD

TRIGGERING OF THE BCR IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL (ten

Hacken, Houston)

CLL INDUCES SEVERE SKEWING IN THE MYELOID COMPARTMENT IN

PATIENTS AND IN THE TCL1 MOUSE MODEL (Hanna, Heidelberg)

Kikushige Y et al., Cancer Cell 2011

HSC in CLL patients generate Clonal B cells

Gene

AA

change Chr CD3+ CD14+

CD5+/

CD19+ CD56+

CD34+/

CD19-

Peripheral

neutrophils

CD19+/CD5-

κ+/λ-

CD19+/CD5-

κ-/λ+

CD34+

derived

colony

frequency

BRAF G469R chr7 11% 24% 50% 29% 8% 14% 48% 9% 51/212 = 24%

NOTCH1 P2515Rfs chr9 - 6% 50% - - - 49% - 5/96 = 5%

DSP T1220I chr6 - - 45% - - - 49% - nd

DMGDH E295K chr5 - - 42% - - - 36% - nd

FBP1 V66M chr9 - - 44% - - - 44% - nd

WDR85 A46V chr9 - - 44% - - - 20% - nd

IRS4 S1091N chrX - - 44% - - - 29% - nd

ODZ1 T1109A chrX - - 47% - - - 30% - nd

SNX8 G306S chr7 - - 19% - - - 14% - nd

ADAD2 C268X chr16 - - 31% - - - 32% - nd

C11orf57 E84K chr11 - - 24% - - - 15% - nd

CCDC113 R127W chr16 - - 22% - - - 21% - nd

KIAA1244 C647S chr6 - - 18% - - - 17% - nd

PCDHA5 S290T chr5 - - 17% - - - 16% - nd

RAG1 R737H chr11 - - 16% - - - 13% - nd

Acquired mutations in HSC of CLL patients

Damm et al., CLL Biology Session EHA 2014

Mutated CD34+ progenitors differentiate into both

lymphoid and myeloid cells

14/24 CLL (58%)

CLL09

0%

10%

20%

30%

40%

50%

60%

0 1 2 3 4 5CD3 CD14 CD34 CD19

Damm et al., CLL Biology Session EHA 2014


CLL and related disorders – Biological

ACQUIRED INITIATING MUTATIONS IN EARLY HEMATOPOIETIC CELLS OF

CHRONIC LYMPHOCYTIC LEUKEMIA PATIENTS (Damm, Villejuif)

SF3B1 MUTATIONS INDUCE COMMON AND LINEAGE SPECIFIC ABERRANT

MESSENGER RNA SPLICING IN MALIGNANCIES INCLUDING CHRONIC

LYMPHOCYTIC LEUKEMIA AND CONFER SENSITIVITY TO SPLICEOSOME

INHIBITION (Buonamici, Cambridge, US)


MODULATION, MICRORNA (MIRNA) REGULATION AND PREDICTION POWER. A

PROSPECTIVE MULTICENTER O-CLL STUDY (Morabito, Cosenza)

ISOTYPE MATTERS: FUNCTIONAL DIFFERENCES AFTER SIGM VERSUS SIGD

TRIGGERING OF THE BCR IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL (ten

Hacken, Houston)



Common events

CLL-specific

Breast-specificMelanoma-specific

SF3B1 mutations lead to aberrant splice events

Buonamici et al., CLL Biology Session EHA 2014

Expression of SF3B1MUT upregulates mutant associated

splice isoforms

a-HA

a-GAPDH

a-SF3B1

a-GAPDH

K6

66

E

K6

66

N

H6

62

Q

K7

00

E

G7

42

D

WT

K7

00

E+R

10

74

H

Vec

tor

con

tro

l

R6

25

C

R6

25

H

E62

2D

SF3B1 cDNA

293FT cells transfected with SF3B1 cDNA

Total RNA extracted for Nanostring

48hr

No

tra

nsf

ecti

on

Expression of WT and MUT isoforms (Nanostring assay)

SF3B1MUT

Buonamici et al., CLL Biology Session EHA 2014


CLL and related disorders: Cinical 1

ABT-199 (GDC-0199) IN RELAPSED/REFRACTORY CHRONIC LYMPHOCYTIC

LEUKEMIA AND SMALL LYMPHOCYTIC LYMPHOMA: HIGH RESPONSE RATES

AMONG PATIENTS WITH HIGH RISK DISEASE FEATURES INCLUDING UNMUTATED

IGHV) (Seymour, Melbourne)

ABT-199 (GDC-0199) COMBINED WITH RITUXIMAB (R) IN PATIENTS (PTS) WITH

RELAPSED / REFRACTORY (R/R) CHRONIC LYMPHOCYTIC LEUKEMIA (CLL):

INTERIM RESULTS OF A PHASE 1B STUDY (Roberts, Melbourne)

SECOND INTERIM ANALYSIS OF A PHASE 3 STUDY EVALUATING IDELALISIB

AND RITUXIMAB FOR RELAPSED CLL (Coutre, Stanford)

A PHASE I STUDY OF THE ORAL BTK INHIBITOR ONO-4059 IN PATIENTS WITH

RELAPSED/REFRACTORY AND HIGH RISK CHRONIC LYMPHOCYTIC LEUKAEMIA

(CLL) (Fegan, Cardiff)

THE ADDITION OF CD20 MONOCLONAL ANTIBODY TO LENALIDOMIDE

IMPROVES RESPONSE RATE AND SURVIVAL IN RELAPSED/REFRACTORY

PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA (Thompson, Houston)









Ulm)










PI3K

p85

CD19

Igb

Iga

PIP3

IP3

Ca++

Sintesi proteica

Trascrizione

DAG

ERCa++

Ca++

Grb2ShcBCAP

PI3K

p100δ

BLNK/SLP65

BtkSyk

Bimp1

MALT1

BCL10

PKCβ

AKT

mTORIKK

PLC-γ2

Calcineurin

NFκB

NFκB

NFAT

NFAT

IκB

Lyn

Fostamatinib

Idelalisib

Ibrutinib

BCR

TLR

IRAK4

IRAK1

TRAF6

TAK1

Randomized Comparison of Ibrutinib Versus Ofatumumab In Previously Treated Chronic Lymphocytic Leukemia / Small Lymphocytic Lymphoma: Results From The Phase Iii Pcyc-1112 Resonate(tm) Trial

#S693 - Presidential Symposium - Saturday 14th June at 15:00 - Room Gold (SW - Level 2)

ted in NEJM1

1. Byrd JC et al. N Engl J Med 2014; DOI:10.1056/NEJMoa1400376

mPFS: not yet reached (ibrutinib) vs. 8.1 months (ofatumumab) (HR 0.22, P<0.001)

6-month PFS (ibrutinib) 88%

OS: HR 0.43, P<0.005

12-month OS: 90% (ibrutinib) vs. 81% (ofatumumab)

ORR: 42.6% (ibrutinib) vs. 4.1% (ofatumumab) (P<0.001)

An additional 20% of ibrutinib-treated patients had PR with lymphocytosis

Similar efficacy in del17p patients and patients with resistance to purine

The most frequent non-hematologic AEs included diarrhea, fatigue, pyrexia, and nausea

Bleeding-related AEs (any grade) occurred in 44% (ibrutinib) vs. 12% (ofatumumab)

23

24

25

26









Ulm)










28

29

30









Ulm)










32

33

34

35









Ulm)










Ibrutinib inhibits NK activation and ADCC at doses reached in vivo

Ibrutinib inhibits phagocytosis of CLL by macrophages in vivo

Ibrutinib inhibits nuetrophils activation and phagocytosis

Comparison between Ibrutinib and Idelalisib

Burger et al, Lancet Oncol 2014

Burger et al, Lancet Oncol 2014


















Study 116: Randomized, Double-Blind, Placebo-Controlled

Idelalisib (150 mg BID)

Placebo (BID)

Rituximab (6 months)

Dis

ease

Pro

gre

ssi

on

Scre

en

Double-BlindInitial Therapy Blinded Dose Open-Label

Extension Study 117



Interim Analyses and Unblinding

Blinded, Independent

Review

Double-BlindContinuous

Therapy

• Primary Endpoint: PFS; Secondary: ORR, LNR, OS

• Interim analyses (IAs) planned at 50% and 75% of total events,

DMC recommended early study stop after 1st IA (Furman et al., NEJM 2014)

• 2nd IA conducted at end of the blinded-phase according to amendment

(data cut-off 09 October 2013 with 63% of total PFS events)

Rituximab (6 months)

Randomizationand Stratification

Primary Study 116

Independent Review

Lymph Node Response

45

-1 0 0

-7 5

-5 0

-2 5

0

2 5

5 0

7 5

1 0 0

1 2 5

Be

st

% C

ha

ng

e i

n S

PD

Id e la lis ib + R itu x im a b

n = 1 0 2a

P la c e b o + R itu x im a b

n = 1 0 1a

aE v a lu a b le p a t ie n t s

D e l(1 7 p ) a n d / o r T P 5 3 m u t

N e it h e r D e l(1 7 p ) o r T P 5 3 m u t

Primary Endpoint: PFS

46

0 2 4 6 8 1 0 1 2 1 4 1 6 1 80

2 0

4 0

6 0

8 0

1 0 0

Pro

gre

ss

ion

-fre

e S

urv

iva

l (%

)

Id e la lis ib + R it u x im a b

M e d ia n P F S : N o t r e a c h e d

P la c e b o + R it u x im a b

M e d ia n P F S : 5 .5 m o n t h s

Id e la lis ib + R 1 1 0 (0 ) 8 7 (3 ) 5 4 (7 ) 3 5 (8 ) 3 0 (1 0 ) 1 7 (1 4 ) 7 (1 5 ) 2 (1 6 ) 1 (1 6 ) 0 (1 6 )

P la c e b o + R 1 1 0 (0 ) 6 9 (2 1 ) 3 7 (3 7 ) 1 9 (4 4 ) 1 4 (4 9 ) 6 (5 4 ) 3 (5 6 ) 1 (5 8 ) 0 (5 9 ) 0 (5 9 )

T im e (M o n th s )N a t r is k (E v e n t s )

H R = 0 .1 8 ,

9 5 % C I (0 .1 0 , 0 .3 2 ),

P < 0 .0 0 0 1

SAE, n (%) IDELA + R (N=110) Placebo + R (N=108)

Patients with any SAE 54 (49) 41 (38)Pneumonia 10 (9) 11 (10)Pyrexia 10 (9) 3 (3)Febrile neutropenia 5 (5) 5 (5)Sepsis 5 (5) 3 (3)Pneumonitis 4 (4) 1 (1)Pneum. jirov. pneumonia 3 (3) 1 (1)Diarrhea 3 (3) 0Hypercalcemia 2 (2) 2 (2)Abdominal pain 2 (2) 1 (1)Hypoxia 2 (2) 1 (1)Colitis 2 (2) 0Deep vein thrombosis 2 (2) 0Sepsis syndrome 2 (2) 0Neutropenia 2 (2) 0Neutropenic sepsis 2 (2) 0Lung infection 2 (2) 0Transient ischemic attack 2 (2) 0

47* Occurring in ≥2 patients on IDELA + R

Serious Adverse Events (SAEs)*

Category, n (%)IDELA + R (N=110) Placebo + R (N=108)

Any Gr. Gr. 3/4 Any Gr. Gr. 3/4

Anemia 32 (29) 8 (7) 35 (32) 18 (17)

Neutropenia 66 (60) 41 (37) 55 (51) 29 (27)

Thrombocytopenia 21 (19) 12 (11) 34 (32) 19 (18)

ALT/AST elevation* 44 (40) 9 (8) 22 (20) 1 (1)

Laboratory Abnormalities of Interest

48

* 7 of the 9 patients in the IDELA + R arm were successfully re-challenged:

• 3 at 150 mg BID, 4 at the reduced dose of 100 mg BID

• One patient’s transaminase elevation was associated with CLL transformation (Richter) in the

liver (progressive disease).









Ulm)










ORR In Subpopulations

50

Favors

Control

Favors

Treatmen

t

* No subjects in the control arm reached the overall response status of CR or PR as of date of analysis

** All subjects in the treatment arm reached the overall response status of CR or PR as of date of analysis

Subgroup OR 95%CI

IDELA+R PLACEBO+R

n ORR(%) n ORR(%)

Overall 29.92 12.76,70.11 88 80.7 88 12.5

del(17p)positive* NA NA,NA 20 80.0 24 0.00

del(17p)negative 21.01 8.46,52.18 68 80.9 64 17.2

TP53mutationpositive 41.51 8.12,212.26 34 79.4 30 10.0

TP53mutationnegative 23.16 8.57,62.60 54 81.5 58 13.8

Bothdel(17p)andTP53mutation* NA NA,NA 17 76.5 15 0.00

Eitherdel(17p)orTP53mutation 48.00 7.23,318.84 20 85.0 24 12.5

Neitherdel(17p)norTP53mutation 20.46 7.31,57.26 51 80.4 49 16.3

del(11q)positive 175.82 11.48,2692.29 28 78.6 29 6.9

del(11q)negative 22.03 8.27,58.69 57 82.5 59 15.3

IGHVmutated 62.92 5.79,683.55 17 88.2 16 12.5

IGHVunmutated 26.36 10.52,66.06 71 78.9 72 12.5

ZAP70positive 28.73 11.33,72.85 77 79.2 75 13.3

ZAP70negative** NA NA,NA 8 100.0 12 8.3

CD38positive 29.27 7.67,111.65 43 83.7 34 17.6

CD38negative 30.32 9.57,96.09 44 77.3 54 9.3

ß2-microglobulin:<=4mg/L** NA NA,NA 12 100.0 18 16.7

ß2-microglobulin:>4mg/L 23.13 9.24,57.89 74 77.0 68 11.8

Del(17p): Best Nodal Response

51

-1 0 0

-7 5

-5 0

-2 5

0

2 5

5 0

7 5

1 0 0

1 2 5

Be

st

% C

ha

ng

e i

n S

PD ID E L A + R it u x im a b P la c e b o + R it u x im a b

N o d e l(1 7 p ) (n = 6 2 )

d e l(1 7 p ) (n = 2 2 )

N o d e l(1 7 p ) (n = 6 7 )

d e l(1 7 p ) (n = 1 8 )

Subgroup HR 95%CI

IDELA+R PLACEBO+R

n PFS n PFS

Overall 0.15 0.08,0.28 110 NR 110 5.5

del(17p)positive 0.14 0.04,0.47 26 NR 31 3.7

del(17p)negative 0.14 0.07,0.31 84 NR 79 6.5

TP53mutationpositive 0.11 0.04,0.31 42 NR 39 3.8

TP53mutationnegative 0.15 0.06,0.37 68 NR 71 7.1

Bothdel(17p)andTP53mutation 0.13 0.04,0.47 22 NR 21 3.5

Eitherdel(17p)orTP53mutation 0.09 0.02,0.42 24 NR 28 5.6

Neitherdel(17p)norTP53mutation 0.17 0.07,0.43 64 NR 61 7.1

del(11q)positive 0.16 0.05,0.59 36 NR 32 6.9

del(11q)negative 0.14 0.07,0.31 71 NR 75 5.5

IGHVmutated 0.25 0.07,0.95 19 12.1 17 11.1

IGHVunmutated 0.13 0.06,0.27 91 NR 93 5.0

ZAP70positive 0.13 0.06,0.25 98 NR 93 5.0

ZAP70negative* NA NA,NA 9 NR 16 8.3

CD38positive 0.13 0.05,0.34 62 NR 51 6.9

CD38negative 0.13 0.05,0.34 47 NR 59 5.5

ß2-microglobulin:<=4mg/L* NA NA,NA 16 NR 24 11.1

ß2-microglobulin:>4mg/L 0.14 0.07,0.27 92 NR 81 3.8

PFS In Subpopulations

52

Favors

Control

Favors

Treatmen

t

* Subjects in the treatment arm are 100% censored as of date of analysis

Del(17p): PFS

Idelalisib

+

Rituxima

b

Placebo

+

Rituxima

b

No 17p- (n=84)

No 17p- (n=79)

17p- (n=26)

17p- (n=31)

Del(17p) and/or TP53 mutations: PFS

54

Idelalisib

+

Rituxima

b

Placebo

+

Rituxima

b

Either 17p- or

TP53mut

(n=24)

Either 17p- or

TP53mut

(n=28)

Both 17p- and

TP53mut (n=22)

Both 17p- and

TP53mut

(n=21)

Del(11q): PFS

55

Idelalisib

+

Rituxima

b

Placebo

+

Rituxima

b

No 11q- (n=71)

No 11q-

(n=75)

11q-

(n=36)

11q-

(n=32)

IGHV Mutation Status: PFS

Idelalisib

+

Rituxima

b

Placebo

+

Rituxima

b

IGHV mut

(n=19)

IGHV mut

(n=17)

IGHV unmut

(n=91)

IGHV unmut (n=93)

β2-Microglobulin: PFS

Idelalisib

+

Rituxima

b

Placebo

+

Rituxima

b

ß2-M ≤ 4 mg/L

(n=16)

ß2-M ≤ 4 mg/L

(n=24)

ß2-M > 4 mg/L

(n=92)

ß2-M > 4 mg/L

(n=81)


















59

61

62

63

64

QUESTIONS

2. How would you choose between the new inhibitors?

3. How would you treat a CLL patients in first line?

1. Can we have easier names for the new drugs?1. Should the new genetic mutations be included in the

work-up before treatment?

CLL: MRD as a Surrogate Endpoint for Clinical Trials

White Oak – February 27, 2013

THE END


Novel CLL genetics – Clinical implications

ACQUIRED INITIATING MUTATIONS IN EARLY HEMATOPOIETIC CELLS

OF CHRONIC LYMPHOCYTIC LEUKEMIA PATIENTS (Damm, Villejuif)

SF3B1 MUTATIONS INDUCE COMMON AND LINEAGE SPECIFIC

ABERRANT MESSENGER RNA SPLICING IN MALIGNANCIES INCLUDING

CHRONIC LYMPHOCYTIC LEUKEMIA AND CONFER SENSITIVITY TO

SPLICEOSOME INHIBITION (Buonamici, Cambridge, US)


MODULATION, MICRORNA (MIRNA) REGULATION AND PREDICTION

POWER. A PROSPECTIVE MULTICENTER O-CLL STUDY (Morabito,

Cosenza)

ISOTYPE MATTERS: FUNCTIONAL DIFFERENCES AFTER SIGM VERSUS

SIGD TRIGGERING OF THE BCR IN CHRONIC LYMPHOCYTIC LEUKEMIA

(CLL (ten Hacken, Houston)



CLL induces T cell abnormalities including an impaired

immunological synapse

Ramsay A. ERIC Symposium EHA 2014; JCI, 2010; Ramsay et al, Blood 2012

T cell

a-CD3/a-CD28

CLL cell

T cell

T cell

T cell

a-CD3/a-CD28

T cell

T cell

T cell proliferation measured by CFSEDilution Assay

Primary human cells

Eµ-TCL1

Suppressive activity of CLL monocytes

CFSE

% o

f m

ax

T cells alone

PBS

Anti-CD3/CD28

T-cells alone

T-cells + monos 1:1

1:2 Monos : T cells

T-cells alone

T-cells +

monos 1:2

1:1 Monos : T cells

Seiffert M, ERIC Symposium, EHA 2014; Hanna et al, CLL Biology Session EHA 2014; Jitschin e al, Blood 2014

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Documents

Paolo Ghia - Ematologialasapienza.it 19 Sett/P. Ghia.pdf · sf3b1 mutations induce common and lineage specific aberrant messenger rna splicing in malignancies including chronic lymphocytic