Human Adenylate Kinase 2 Deficiency Causes a Profound Haematopoietic Defect Associated with Sensorineural Deafness

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. lba-2-lba-2
Author(s):  
Marina Cavazzana-Calvo ◽  
Chantal Lagresle-Peyrou ◽  
Emmanuelle M Six ◽  
Capucine Picard ◽  
Frederic Rieux-Laucat ◽  
...  
Keyword(s):  
Adenylate Kinase ◽  
Bone Marrow Cells ◽  
Stria Vascularis ◽  
Severe Combined Immunodeficiency ◽  
Sensorineural Deafness ◽  
Polymorphonuclear Neutrophils ◽  
Recessive Form ◽  
Severe Infections ◽  
Differentiation Block ◽  
Neutrophil Differentiation

Abstract Human adenylate kinase 2 deficiency causes the most profound human haematopoietic defect associated with sensorineural deafness. Reticular dysgenesis (RD) is an autosomal recessive form of human severe combined immunodeficiency (SCID) characterized by an early differentiation block in the myeloid lineage and a profound impairment in lymphoid maturation associated with bilateral sensorineural deafness. The lack of polymorphonuclear neutrophils in affected newborns is responsible for the occurrence of severe infections earlier than usually observed in the other forms of SCID. Furthermore, RD associated neutropenia is characterized by the lack of responsiveness to G-CSF. We have identified bi-allelic mutations in the adenylate kinase 2 (AK2) gene is seven patients affected with RD. These mutations resulted in the absence or a strong decrease in protein expression. Restoration of AK2 expression in the bone marrow cells of RD patients overcomes the neutrophil differentiation arrest, underlying its specific requirement in the development of a restricted set of haematopoietic lineages. Lastly, we established that AK2 is specifically expressed in the stria vascularis region of the inner ear. The function of this gene in the differentiation of a given set of cell lineages is rapidly in progress, showing that studies of primary immunodeficiencies continue to provide key information on human lympho-haematopoietic development. Moreover the AK2 enzyme seems a key molecule in different biological systems such as the lympho-haematopoiesis and the brain development.

Human Adenylate Kinase 2 Deficiency Inhibits Hematopoietic Cell Differentiation towards Neutrophil and T Lymphoid Lineages.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 78-78
Author(s):  
Emmanuelle M. Six ◽  
Chantal Lagresle-Peyrou ◽  
Corinne Demerens-deChappedelaine ◽  
Frederic Rieux-Laucat ◽  
Andrea Didati ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Cell Line ◽  
Adenylate Kinase ◽  
Conflicts Of Interest ◽  
Adenine Nucleotides ◽  
Severe Combined Immunodeficiency ◽  
Mitochondrial Protein ◽  
Hematopoietic Cell ◽  
Hematopoietic Stem ◽  
Neutrophil Differentiation

Abstract Abstract 78 Reticular dysgenesis (RD), an autosomal recessive form of human Severe Combined Immunodeficiency is characterized by the absence of blood neutrophils and T lymphocytes. This pathology is due to biallelic mutations in the adenylate kinase 2 (AK2) gene, resulting in the loss of AK2 protein expression. AK2 is a mitochondrial protein which regulates the homeostasis of cellular adenine nucleotides by converting ADP into ATP and AMP. In order to understand the precise role of AK2 in hematopoiesis, we have developed a RNA interference strategy through lentiviral-mediated gene transfer of AK2 short hairpin RNAs (shAK2). The knock-down of AK2 in human or murine hematopoietic stem cells (HSC) inhibits their capacity to form granulocyte colonies in methycellulose assays and prevents them to generate mature polynucleated cells in liquid culture in the presence of G-CSF. We also determine the ability of shAK2-transduced HSC to differentiate along the T lymphoid lineage after co-culture on a OP9Delta1 stroma cell line. Our data demonstrated that the apparition of CD4+CD8+ cells was profoundly reduced in the presence of shAK2. To delineate the mechanism involved in this defect, we also studied the neutrophil differentiation of the HL60 promyelocytic cell line, following retinoic acid treatment. In this system, the absence of AK2 expression led to an arrest of neutrophil differentiation process, increased cell apoptosis and disrupt the mitochondrial membrane potential. All these data suggest a novel mechanism regulating hematopoietic cell differentiation, and involved in one of the most severe human immunodeficiency syndromes. Disclosures: No relevant conflicts of interest to declare.

Adenylate Kinase 2 deficiency causes NAD+ depletion and impaired purine metabolism during myelopoiesis

2021 ◽  
Author(s):  
Wenqing Wang ◽  
Andrew DeVilbiss ◽  
Martin Arreola ◽  
Thomas Mathews ◽  
Misty Martin-Sandoval ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Adenylate Kinase ◽  
De Novo ◽  
Bone Marrow Cells ◽  
Severe Combined Immunodeficiency ◽  
Purine Metabolism ◽  
Model Systems ◽  
Hematopoietic Stem ◽  
Mechanistic Basis ◽  
Nad Depletion

Reticular Dysgenesis is a particularly grave from of severe combined immunodeficiency (SCID) that presents with severe congenital neutropenia and a maturation arrest of most cells of the lymphoid lineage. The disease is caused by biallelic loss of function mutations in the mitochondrial enzyme Adenylate Kinase 2 (AK2). AK2 mediates the phosphorylation of adenosine monophosphate (AMP) to adenosine diphosphate (ADP) as substrate for adenosine triphosphate (ATP) synthesis in the mitochondria. Accordingly, it has long been hypothesized that a decline in OXPHOS metabolism is the driver of the disease. The mechanistic basis for Reticular Dysgenesis, however, remained incompletely understood, largely due to lack of appropriate model systems to phenocopy the human disease. We have used single cell RNA-sequencing of bone marrow cells from 2 reticular dysgenesis patients to gain insight into the disease pathology. Gene set enrichment for differentially expressed genes in different subsets of myeloid and lymphoid progenitor cells pointed to processes involving RNA and ribonucleoprotein assembly and catabolism as well as cell cycle defects. To investigate these findings and precisely mimic the failure of human myelopoiesis in culture, we developed a cell-tracible model of Reticular Dysgenesis based on CRISPR-mediated disruption of the AK2 gene in primary human hematopoietic stem cells. In this model, we have identified that AK2-deficienct myeloid progenitor cells exhibit NAD+ depletion and high levels of reductive stress accompanied by an accumulation of AMP and IMP while ADP and ATP are only mildly decreased. Our studies further show that AK2-deficienct cells have decreased de novo purine synthesis and increased purine breakdown, accompanied by decreased RNA and ribosome subunit cellular content. These data highlight the profound impact of mitochondrial dysfunction on the cellular redox state and nucleotide pool and identify the mechanistic basis of Reticular Dysgenesis as a defect in purine metabolism.

ZAP-70 deficiency in an autosomal recessive form of severe combined immunodeficiency

Science ◽  
1994 ◽  
Vol 264 (5165) ◽  
pp. 1599-1601 ◽  
Author(s):  
A. Chan ◽  
T. Kadlecek ◽  
M. Elder ◽  
A. Filipovich ◽  
W. Kuo ◽  
...  
Keyword(s):  
Autosomal Recessive ◽  
Severe Combined Immunodeficiency ◽  
Combined Immunodeficiency ◽  
Autosomal Recessive Form ◽  
Recessive Form

scholarly journals A self-inactivating lentiviral vector for SCID-X1 gene therapy that does not activate LMO2 expression in human T cells

Blood ◽  
2010 ◽  
Vol 116 (6) ◽  
pp. 900-908 ◽  
Author(s):  
Sheng Zhou ◽  
Disha Mody ◽  
Suk See DeRavin ◽  
Julia Hauer ◽  
Taihe Lu ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Bone Marrow ◽  
Protein Expression ◽  
Genomic Sequence ◽  
Bone Marrow Cells ◽  
Lentiviral Vector ◽  
Severe Combined Immunodeficiency ◽  
Vitro Assay ◽  
Activation Assay ◽  
Marrow Cells

Abstract To develop safer and more effective vectors for gene therapy of X-linked severe combined immunodeficiency (SCID-X1), we have evaluated new self-inactivating lentiviral vectors based on the HIV virus. The CL20i4-hγc-Revgen vector contains the entire human common γ chain (γc) genomic sequence driven by the γc promoter. The CL20i4-EF1α-hγcOPT vector uses a promoter fragment from the eukaryotic elongation factor alpha (EF1α) gene to express a codon-optimized human γc cDNA. Both vectors contain a 400-bp insulator fragment from the chicken β-globin locus within the self-inactivating long-terminal repeat. Transduction of bone marrow cells using either of these vectors restored T, B, and natural killer lymphocyte development and function in a mouse SCID-X1 transplantation model. Transduction of human CD34+ bone marrow cells from SCID-X1 patients with either vector restored T-cell development in an in vitro assay. In safety studies using a Jurkat LMO2 activation assay, only the CL20i4-EF1α-hγcOPT vector lacked the ability to transactivate LMO2 protein expression, whereas the CL20i4-hγc-Revgen vector significantly activated LMO2 protein expression. In addition, the CL20i4-EF1α-hγcOPT vector has not caused any tumors in transplanted mice. We conclude that the CL20i4-EF1α-hγcOPT vector may be suitable for testing in a clinical trial based on these preclinical demonstrations of efficacy and safety.

AK2 Deficiency In Zebrafish Recapitulates Human Reticular Dysgenesis, An Autosomal Recessive Form Of Severe Combined Immunodeficiency

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2416-2416
Author(s):  
Alberto Rissone ◽  
Jaya Jagadeesh ◽  
Karen Simon ◽  
Kevin Bishop ◽  
Raman B. Sood ◽  
...  
Keyword(s):  
Autosomal Recessive ◽  
Molecular Mechanisms ◽  
Conflicts Of Interest ◽  
Severe Combined Immunodeficiency ◽  
Combined Immunodeficiency ◽  
Zebrafish Model ◽  
Hematopoietic Stem ◽  
Autosomal Recessive Form ◽  
Recessive Form ◽  
Mutant Lines

Abstract The adenylate kinase (AK) gene family consists of 7 different members that contribute to energy cell metabolism by converting ATP+AMP to 2ADP. AKs are critical players in ensuring cellular energy homeostasis in all tissues. Mutations in the AK2 gene are responsible for reticular dysgenesis (RD), an autosomal recessive form of severe combined immunodeficiency (SCID). RD is characterized by an early differentiation arrest in the granulocyte lineage and impaired lymphoid maturation and it represents less than 2% of total SCID. Affected children succumb to overwhelming infections early in life unless their immune system is successfully restored with allogeneic hematopoietic stem cells transplant (HSCT). The mechanisms underlying the pathophysiology of RD remain unclear. The phenotype of AK2 deficient animals has never been reported in the literature, but murine lines carrying homozygous inactivating retroviral insertions are embryonically lethal (our personal observations). We used the zebrafish model to perform a comprehensive study of the effects of AK2 deficiency using Morpholino oligomers injections and two different kinds of AK2 mutants (a ENU-induced T371C/L124P missense mutant and two null mutant lines generated using zinc-finger nuclease technology). In situ hybridization analyses of AK2-deficient embryos indicated that only erythroid development was affected during primitive hematopoiesis. Conversely, during definitive hematopoiesis, the loss of function of AK2 resulted in abnormalities distributed along all hematopoietic lineages suggesting an impairment of hematopoietic stem cell (HSC) development. Moreover, we observed that the AK2 deficiency induced oxidative stress and consequent apoptosis in both primitive erythroid cells and definitive HSCs. Importantly, antioxidant treatment of AK2 mutant embryos rescued the hematopoietic phenotypes as indicated by the recovered expression of HSC and lymphoid markers (such as c-myb and rag1). Overall, our data indicate that zebrafish represents a good model for studying the molecular mechanisms involved in RD and testing of new therapeutic interventions. To date, our mutant lines remain the only animal model of this rare and lethal human disease. Disclosures: No relevant conflicts of interest to declare.

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