NRAS mutation causes a human autoimmune lymphoproliferative syndrome

João Oliveira

Nicolas Bidère

Julie Niemela

Lixin Zheng

Peter Munson+5 authors

*Department of Laboratory Medicine, Clinical Center,

^{Molecular Development Section, Laboratory of Immunology, National Institute of Allergy and Infectious Diseases,}

^{Functional Genomics and Proteomics Facility, Critical Care Medicine Department, Clinical Center,}

^{Mathematical and Statistical Computing Laboratory, Center for Information Technology,}

**Genetics and Molecular Biology Branch, National Human Genome Research Institute, and

^{Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892}

^{To whom correspondence may be addressed at: Clinical Center, National Institutes of Health, MSC 1508, Bethesda, MD 20892., E-mail:}vog.hin.liam@ehsielft

^{To whom correspondence may be addressed at: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, MSC 1892, Bethesda, MD 20892., E-mail:}vog.hin.diain@odranelm

Communicated by Jacques F. A. P. Miller, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia, April 3, 2007.

Author contributions: J.B.O., T.A.F., and M.J.L. designed research; J.B.O., N.B., J.E.N., L.Z., K.S., and C.P.N. performed research; R.L.D. and J.M.P. contributed new reagents/analytic tools; J.B.O., N.B., J.E.N., R.L.D., J.B., P.J.M., T.A.F., and M.J.L. analyzed data; J.B.O., T.A.F., and M.J.L. wrote the paper; J.D. coordinated patient information and clinical sample handling; and S.E.S. coordinated the clinical aspects of the research.

^{Present address: Laboratory of Medical Investigation (LIM-56), University of São Paulo, 05630-020, São Paulo, Brazil.}

^{Present address: Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL 33136.}

Received 2007 Jan 5

Abstract

The p21 RAS subfamily of small GTPases, including KRAS, HRAS, and NRAS, regulates cell proliferation, cytoskeletal organization, and other signaling networks, and is the most frequent target of activating mutations in cancer. Activating germline mutations of KRAS and HRAS cause severe developmental abnormalities leading to Noonan, cardio-facial-cutaneous, and Costello syndrome, but activating germline mutations of NRAS have not been reported. Autoimmune lymphoproliferative syndrome (ALPS) is the most common genetic disease of lymphocyte apoptosis and causes autoimmunity as well as excessive lymphocyte accumulation, particularly of CD4^{, CD8}^{αβ T cells. Mutations in ALPS typically affect CD95 (Fas/APO-1)-mediated apoptosis, one of the extrinsic death pathways involving TNF receptor superfamily proteins, but certain ALPS individuals have no such mutations. We show here that the salient features of ALPS as well as a predisposition to hematological malignancies can be caused by a heterozygous germline Gly13Asp activating mutation of the}NRAS oncogene that does not impair CD95-mediated apoptosis. The increase in active, GTP-bound NRAS augments RAF/MEK/ERK signaling, which markedly decreases the proapoptotic protein BIM and attenuates intrinsic, nonreceptor-mediated mitochondrial apoptosis. Thus, germline activating mutations in NRAS differ from other p21 Ras oncoproteins by causing selective immune abnormalities without general developmental defects. Our observations on the effects of NRAS activation indicate that RAS-inactivating drugs, such as farnesyltransferase inhibitors should be examined in human autoimmune and lymphocyte homeostasis disorders.

Keywords: autoimmunity, B cell lymphoma 2-interacting mediator of cell death, intrinsic apoptosis, lymphoma, lymphoproliferation

Abstract

The RAS genes (NRAS, KRAS, and HRAS) encode 21-kDa proteins that are members of the superfamily of small GTP-binding proteins, which have diverse intracellular signaling functions including control of cell proliferation, growth, and apoptosis (1). Somatic activating mutations in RAS are present in up to 30% of all human cancers (2). Germline RAS pathway mutations have only recently been described as causing the related Costello (HRAS), Noonan (PTPN11, KRAS, SOS1), and cardiofaciocutaneous syndromes (KRAS, BRAF, MEK1, and MEK) (3 –7). Individuals with these syndromes typically present with severe developmental anomalies in various combinations of facial abnormalities, heart defects, short stature, skin and genital abnormalities, and mental retardation (8). Defects in the immune system have not been reported. Patients with Costello and Noonan syndromes have an increased propensity to solid and hematopoietic tumors, respectively (3, 8). Germline mutations in NRAS have not yet been described.

The autoimmune lymphoproliferative syndrome (ALPS) (OMIM 601859/603909) is the most common genetic disorder of lymphocyte apoptosis and is characterized by chronic accumulation of nonmalignant lymphocytes, defective lymphocyte apoptosis, and an increased risk for the development of hematological malignancies (9). A signature of the disease is the accumulation of αβ T cells lacking the CD4 and CD8 coreceptors that are termed “double-negative” T cells (DNTs: CD4^{, CD8}^TCRαβ^{cells). These cells bear no known relationship to thymic DNTs, a stage that occurs before αβ TCR gene rearrangements in ontogeny (}10). According to genotype, ALPS can be classified as types Ia, Ib, and II, which are due to germline mutations in CD95 (TNFRSF6), CD95 ligand (TNFSF6), and caspase 10 (CASP10), respectively (10 –13). Additionally, somatic mutations of CD95 in αβ DNTs can also cause ALPS of type Im (mosaic) (14). All of these mutations impair extrinsic, Fas receptor-mediated apoptosis (10). An enigma has been the ALPS individuals who have no defects in CD95 pathway apoptosis (some ALPS type III patients) (9, 10). This group encompasses a large number of individuals and is probably genetically heterogeneous. In an attempt to unveil new genetic defects, we investigated alternative apoptosis pathways in ALPS type III and identified one ALPS patient with a unique defect in cytokine withdrawal-induced apoptosis due to an activating NRAS mutation.

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Acknowledgments

We thank C. Logun for technical assistance; T. Inaba (Hiroshima University, Hiroshima, Japan), J. Gavard (National Institutes of Health), and S. Gutkind for reagents; R. Germain, R. Siegel, and A. Snow for critical reading of the manuscript; and S. Patel for help with cytokine measurement. Cynthia P. Nix was supported by the Howard Hughes Medical Institute–NIH Research Scholar Program. This work was supported by the National Institute of Allergy and Infectious Diseases NIH Intramural Research Program.

Acknowledgments

Abbreviations

ALPS	autoimmune lymphoproliferative syndrome
DNT	double-negative T cell
BCL-2	B cell lymphoma 2
BIM	BCL-2-interacting mediator of cell death
siRNA	small interfering RNA
FTI	farnesyltransferase inhibitor
PBL	peripheral blood lymphocyte.

Abbreviations

Footnotes

The authors declare no conflict of interest.

Data deposition: The data reported in this paper have been deposited in the Gene Expression Omnibus (GEO) database, www.ncbi.nlm.nih.gov/geo (accession no. {"type":"entrez-geo","attrs":{"text":"GSE7345","term_id":"7345","extlink":"1"}}GSE7345).

This article contains supporting information online at www.pnas.org/cgi/content/full/0702975104/DC1.

Footnotes

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