BCL2 family regulatory networks in the context of Bcell lymphomagenesis

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Slide 1 :

BCL2 family regulatory networks in the context of B-cell lymphomagenesis (Antonio Ruiz-Vela, PhD) Theophilius Schweighart, 1604

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Pro-B cell B220+ IgM- CD23- ckit+ CD43+ CD24+ CD23- Schematic representation of B-cell development Pre-BII V-DJ/IgH l5 Pre-BCR Early Pre-BI CD25+ D-J/IgH Pre-BI CD19+ Immature B cell V-J/IgL BCR (IgM+) Autoantigens Programmed Cell Death ‘Negative Selection checkpoint’ (BCL2 family members keep in check) Bone Marrow Spleen Lymph nodes Transitional B cell IgD+ IgM+ CD24+ CD23- CD21- Mature B cell IgD++ IgM+ CD24- CD23+ CD21+ HSC Sca-1+ Lin- ckit+ EBF E2A PAX5 IL7R/IL7

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Bcl-2 Bcl-xL Bcl-w Mcl-1 A1/Bfl-1 Boo/Diva Nr13 BH4 BH3 BH1 BH2 TM Antiapoptotic members BCL2 family members (BCL2 family members are critical regulators of programmed cell death) TM Bax Bak Bok/Mtd Bid Bad Bik/Nbk Blk Hrk Bim/Bod Bnip3 Nix Noxa BH3 BH1 BH2 Proapoptotic members BH3-only proteins Multidomain proteins

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Remodeling of the Mitochondrial Cristae and Cytochrome c release during programmed cell death Scorrano L., Dev Cell. 2002 Jun;2(1):55-67. IM boundary Outer membrane Cristae Class I Class II Class III Acehan D., Mol Cell. 2002 Feb;9(2):423-32. Liu et al., Cell. 1996 Jul 12;86(1):147-57

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Programmed cell death pathway initiated by B-cell receptor (BCR) regulatory networks in B-cells (BAX, BAK) APAF-1/ CASP-9 CAPN-1, -2 Ruiz-Vela et al., Blood. 2008 Feb 1;111(3):1665-76. Ruiz-Vela et al., EMBO J. 1999 Sep 15;18(18):4988-98. Ruiz-Vela et al., J Exp Med. 2001 Aug 6;194(3):247-54. Takeuchi et al., Proc Natl Acad Sci U S A. 2005 Aug 9;102(32):11272-7 ??? ??? BCR Autoantigen

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Combined inactivation of proapoptotic BAX and BAK generates an autoreactive B-cell lymphomagenesis Lindsten., Mol Cell. 2000 Dec;6(6):1389-99. BAX-/-, BAK-/- BAX-/-, BAK+/- BAX+/-, BAK-/- BAX+/-, BAK+/- Lymph node Spleen BAX, BAK BAX, BAK BAX, BAK BAX, BAK

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(BAX, BAK) APAF-1/ CASP-9 CAPN-1, -2 ??? ??? BCR ? Experimental system to dissect key molecules linking BCR signalling and programmed cell death Burkkit’s lymphoma (BL) hallmark IgH enhancer MYC t (8:14) ER membrane Loss-of-function strategy (RNAi)

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Lentiviral (HIV)-based RNA interference screen EV #1 #2 #3

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Methodology in our RNA interference screen

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RNA interference screen in the context of BCR signalling

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MCL1 gain-of-function

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MCL1 interactome map

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Gene network reconstruction analysis

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Acknowledgment: Stanley Korsmeyer Lab Scott Armstrong Joe Opferman

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Acknowledgment: Miguel Angel Piris Lab Beatriz Herreros Daniel Martin Pierfrancesco Vargiu Abel Sanchez-Aguilera

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