Nerve gene expression patterning in Nf1 mouse models

844 views

Published on

Platform presentation at the Children’s Tumor Foundation (CTF) Annual NF Conference 2009, Portland, OR. Session:Novel Cellular and Animal Models of NF.

Published in: Health & Medicine, Technology
0 Comments
0 Likes
Statistics
Notes
  • Be the first to comment

  • Be the first to like this

No Downloads
Views
Total views
844
On SlideShare
0
From Embeds
0
Number of Embeds
11
Actions
Shares
0
Downloads
4
Comments
0
Likes
0
Embeds 0
No embeds

No notes for slide
  • Nerve gene expression patterning in Nf1 mouse models

    1. 1. Nerve gene expression patterning in Nf1 mouse models Walter J. Jessen, Ph.D. Laboratory of Nancy Ratner Division of Experimental Hematology and Cancer Biology Cincinnati Children’s Hospital Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 1
    2. 2. Myelinated vs. unmyelinated axons A. Myelinated Axon B. Unmyelinated Axons Ax S S S Schwann cell nucleus Ax Axon Ax Corfas et al., 2004 Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 2
    3. 3. Mast cell infiltration and Remak bundle disruption  One hallmark of neurofibromatosis type 1 (NF1): infiltration of mast cells in peripheral nerve tumors (Pineda, 1965; Olsson, 1971).  Many mouse models of NF1 display abnormalities in the organization of Remak bundles (Ling et al, 2005; Wu et al., 2008; Zheng et al., 2008).  Coincident with mast cell infiltration is the disruption of Remak bundles and the accumulation of collagen (Monk et al. 2007). Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 3
    4. 4. Models for NF1 tumor formation 1. Linear pathway Normal nerve Disrupted nerve Tumor 2. Mutually exclusive pathway Disrupted nerve Normal nerve Tumor Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 4
    5. 5. Models for NF1 tumor formation 1. Linear pathway Normal nerve Disrupted nerve Neurofibroma 2. Mutually exclusive pathway Disrupted nerve Normal nerve Tumor Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 5
    6. 6. Disruption of Remak bundles in peripheral nerves of EGFR/+ mice correlates with mast cell infiltration Disrupted Remak bundle Monk et al., 2007 Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 6
    7. 7. Disruption of Remak bundles in peripheral nerves of EGFR/+ mice correlates with mast cell infiltration Disrupted Remak bundle Evaluate the acute effects Monk et al., 2007 of mast cells on nerve. 4 Week 8 Week Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 7
    8. 8. The disrupted Remak bundle phenotype is observed in both EGFR over-expression and GEM Nf1 models Wild type EGFR/+ Monk et al., 2007 Monk et al., 2007 Nf1flox/-; P0A-cre+ DhhCre; Nf1 fl/fl Zheng et al., 2008 Wu et al., 2008 Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 8
    9. 9. Objectives Test the hypotheses that genetically engineered mouse (GEM) models showing mast cell infiltration and Remak bundle disruption: 1. Share common nerve gene expression programs and have similar transcription profiles. 2. Maintain nerve gene transcription profiles in GEM-neurofibroma and/or -PNST models. ~100 mouse GEM model samples encompassing 20 genotypes Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 9
    10. 10. Data analysis Nerve gene expression patterns from GEM models of NF1 Nerve gene patterns across GEM model tumors Integrate data sets Identify mouse Identify nerve nerve gene genes changed orthologs that are with early Remak also changed bundle disruption in human and mast cell NF1-derived infiltration Schwann cells Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 10
    11. 11. Nerve gene expression patterns from GEM models of NF1 Hypothesis #1 GEM models showing Nerve gene patterns across mast cell infiltration and GEM model tumors Remak bundle disruption Integrate data sets share common nerve gene expression programs and Identify nerve Identify mouse nerve gene have similar transcription genes changed orthologs that are with early Remak profiles. bundle disruption also changed in human and mast cell NF1-derived infiltration Schwann cells Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 11
    12. 12. Five GEM models of Nf1 and EGFR over-expression known to cause mast cell infiltration and Remak bundle disruption GEM Nf1 models Unknown P0CreB; Nf1 flox/- H-Ras V12 P0CreB; Nf1 flox/flox NPcis (Nf1+/-; p53+/-) DhhCre; Nf1 flox/flox GEM models over-expressing EGFR CNPase-hEGFR/+ CNPase-hEGFR/hEGFR Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 12
    13. 13. GEM model nerves: gene expression profiles Control nerves GEM model nerves C1 C2 4.00 2,028 transcripts C3 C4 0.25 C5 C6 10 genotypes Disrupted Remak bundle phenotype Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 13
    14. 14. GEM model nerves: gene expression profiles Control nerves GEM model nerves C1 C2 4.00 2,028 transcripts C3 C4 0.25 C5 C6 10 genotypes Disrupted Remak bundle phenotype Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 14
    15. 15. GEM models showing mast cell infiltration and Remak bundle disruption share common gene expression programs and have similar transcription profiles GEM model nerves  Microtubule cytoskeleton organization and biogenesis C1  Lipid metabolic process  Integral to membrane (32%) C2 Enrichment p-value < 0.05 4.00 2,028 transcripts  Immune response C3  Chemotaxis C4  Angiogenesis 0.25  Extracellular matrix organization C5 and biogenesis  Wnt signaling pathway  Cell differentiation C6  EGFR signaling pathway  Extracellular region (28%)  Collagen (2%) Disrupted Remak bundle phenotype Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 15
    16. 16. H-Ras and NPcis nerves do not have a disrupted Remak bundle phenotype H-Ras V12 nerve NPcis nerve ** mice do not form tumors ** mice only form PNST Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 16
    17. 17. Nerve gene expression patterns from GEM models of NF1 Hypothesis #2 GEM models showing Nerve gene patterns across mast cell infiltration and GEM model tumors Remak bundle disruption Integrate data sets maintain nerve gene expression profiles in GEM- Identify nerve Identify mouse nerve gene neurofibroma and/or -PNST genes changed orthologs that are with early Remak models. bundle disruption also changed in human and mast cell NF1-derived infiltration Schwann cells Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 17
    18. 18. Nf1 nerves: Gene expression profiles in GEM tumors Mouse nerves Mouse tumors Control nerves Control Nerves GEM model nerves GEM-Neurofibroma GEM-PNST GEM-neurofibroma C1 C2 4.00 2,028 transcripts C3 0.25 C4 C5 C6 20 genotypes Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 18
    19. 19. GEM models showing mast cell infiltration and Remak bundle disruption Nf1 nerves: Genetranscription profiles in GEM-Neurofibroma and -PNST maintain nerve gene expression profiles in GEM tumors Mouse nerves Mouse tumors Control nerves Control Nerves GEM model nerves GEM-Neurofibroma GEM-PNST GEM-neurofibroma C1 C2 4.00 2,028 transcripts C3 0.25 C4 C5 C6 20 genotypes Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 14 19
    20. 20. Early features of nerve disruption Evaluate the acute effects of Nerve gene expression patterns mast cells on nerve. from GEM models of NF1 Strategy: Nerve gene patterns across use GEM Nf1 and EGFR GEM model tumors over-expression models to compare nerve gene Integrate data sets programs expressed at Identify mouse 4-weeks and 8-weeks Identify nerve nerve gene genes changed orthologs that are with early Remak also changed bundle disruption in human and mast cell NF1-derived infiltration Schwann cells Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 20
    21. 21. With early Remak bundle disruption and mast cell infiltration, there is a clear difference in gene patterning between controls and GEM model nerves Genes also changed 4 week-old nerve 8 week-old nerve in nerve gene clusters C1 and C6 4.00 1339 transcripts 0.25 3 genotypes Nf1 flox/flox EGFR/EGFR DhhCre; Nf1 flox/flox Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 21
    22. 22. Nerve differentiation and maturation gene programs are lost in EGFR/EGFR and DhhCre;Nf1 flox/flox nerves Genes also changed 4 week-old nerve 8 week-old nerve in nerve gene clusters C1 and C6 4.00 1339 transcripts 0.25 3 genotypes Nf1 flox/flox EGFR/EGFR DhhCre; Nf1 flox/flox Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 22
    23. 23. Failed nerve maturation correlates with gene signatures of Remak bundle disruption in GEM Nf1 and EGFR nerve gene expression Genes also changed 4 week-old nerve 8 week-old nerve in nerve gene clusters C1 and C6 4.00 1339 transcripts 0.25 3 genotypes Nf1 flox/flox EGFR/EGFR DhhCre; Nf1 flox/flox Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 23
    24. 24. Schwann cell component Nerve gene expression patterns from GEM models of NF1 Which mouse nerve genes are also changed in human NF1-derived Schwann cells? Nerve gene patterns across GEM model tumors Integrate data sets Strategy: compare mouse nerve gene Identify nerve Identify mouse orthologs from clusters C1 nerve gene genes changed orthologs that are and C6 to NF1-derived with early Remak also changed bundle disruption Schwann cells and primary in human and mast cell NF1-derived tumor gene programs. infiltration Schwann cells Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 24
    25. 25. GEM disrupted nerve genes that are also differentially expressed in NF1-derived Schwann cells (SCs) and MPNST cell lines Genes also in mouse Human Schwann cell cultures Primary human tumors nerve gene clusters C1 and C6 6.00 268 Gene Orthologs 0.00 * * * MPNST NHSC NF1-derived SCs Neurofibroma MPNST cell lines Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 25
    26. 26. Summary  GEM models showing mast cell infiltration and Remak bundle disruption share common nerve gene expression programs.  GEM models showing mast cell infiltration and Remak bundle disruption maintain nerve gene transcription profiles in GEM- neurofibroma and GEM-PNST.  Nerve differentiation/maturation is lost in GEM Nf1 and EGFR over-expression models.  Failed nerve differentiation/maturation correlates with Remak bundle disruption.  Abnormal Schwann cell signatures are associated with Remak bundle disruption in GEM model nerves, GEM-neurofibromas, GEM-PNST and human neurofibromas and MPNST. Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 26
    27. 27. Mouse models Human Cincinnati, OH – CCHMC Cincinnati, OH – CCHMC Nancy Ratner, Jianqiang Wu, Tilat Rizvi Nancy Ratner, Shrya Miller, Atira Hardiman Paris, France – Fondation Jean Dausset Boston, MA – MGH/Harvard Marco Giovannini, Jan Manent Anat Stemmer-Rachamimov Gainseville, FL – University of Florida Bioinformatics/Biostatistics Margaret Wallace Cincinnati, OH – CCHMC Bruce Aronow, Walter Jessen Barcelona, Spain – LʼHospitalet de Llobregat Concepcion Lazaro, Eduard Serra Birmingham, AL – University of Alabama Grier Page, Tapan Mehta Funding DOD W81XWH-04-1-0273 Walter Jessen Nf1 mouse nerve gene expression Walter.Jessen@cchmc.org 27

    ×