Andreas Holzinger

Technische Universität München, München, Bavaria, Germany

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Publications (66)78.31 Total impact

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    ABSTRACT: Background: Heterozygous ATP-binding-cassette subfamily A member 3 (ABCA3) mutations are associated with neonatal respiratory complications. In an adult murine model, we investigated whether Abca3 haploinsufficiency is a predisposing factor for lung injury induced by hyperoxia or mechanical ventilation. Methods: Abca3 haploinsufficient (Abca3(+/-)) and wild-type (WT) mice were prospectively randomized to 25 min of ventilation or 72 h of hyperoxia or left unchallenged in air. Results: As compared with WT mice, unchallenged Abca3(+/-) mice had significantly decreased lung phosphatidylcholine (PC) and phosphatidylglycerol (PG) levels (P < 0.02) and decreased lung compliance (P < 0.05). When ventilated for 25 min, Abca3(+/-) mice demonstrated a significantly greater increase in bronchoalveolar lavage (BAL) interleukins (P ≤ 0.01) and lung wet to dry ratio (P < 0.005). Hyperoxia resulted in increased compliance (P < 0.05) and total lung capacity (TLC) (P = 0.01) only in the Abca3(+/-) mice, consistent with enlarged alveolar spaces. The ratio of PC to PG in BAL-relevant for surfactant dysfunction-was significantly elevated by oxygen exposure, with the greatest increase in Abca3(+/-) mice. Conclusion: In a murine model, Abca3 haploinsufficiency results in an altered biochemical and lung mechanical phenotype, as well as a greater lung injury induced by hyperoxia or mechanical ventilation. The inability to maintain a normal PC/PG ratio appears to play a key role.
    Pediatric Research 07/2013; 74(4). DOI:10.1038/pr.2013.127 · 2.31 Impact Factor
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    ABSTRACT: Tissue-specific transcripts are likely to be of importance for the corresponding organ. While attempting to define the specific transcriptome of the human lung, we identified the transcript of a yet uncharacterized protein, SFTA2. In silico analyses, biochemical methods, fluorescence imaging and animal challenge experiments were employed to characterize SFTA2. Human SFTA2 is located on Chr. 6p21.33, a disease-susceptibility locus for diffuse panbronchiolitis. RT-PCR verified the abundance of SFTA2-specific transcripts in human and mouse lung. SFTA2 is synthesized as a hydrophilic precursor releasing a 59 amino acid mature peptide after cleavage of an N-terminal secretory signal. SFTA2 has no recognizable homology to other proteins while orthologues are present in all mammals. SFTA2 is a glycosylated protein and specifically expressed in nonciliated bronchiolar epithelium and type II pneumocytes. In accordance with other hydrophilic surfactant proteins, SFTA2 did not colocalize with lamellar bodies but colocalized with golgin97 and clathrin-labelled vesicles, suggesting a classical secretory pathway for its expression and secretion. In the mouse lung, Sfta2 was significantly downregulated after induction of an inflammatory reaction by intratracheal lipopolysaccharides paralleling surfactant proteins B and C but not D. Hyperoxia, however, did not alter SFTA2 mRNA levels. We have characterized SFTA2 and present it as a novel unique secretory peptide highly expressed in the lung.
    PLoS ONE 06/2012; 7(6):e40011. DOI:10.1371/journal.pone.0040011 · 3.23 Impact Factor
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    ABSTRACT: Tissue-specific transcripts are likely to play a significant role in the function of the corresponding organ. In an in silico approach to define the specific transcriptome of the human lung, we identified the transcript of a yet uncharacterized protein, SFTA2. The human SFTA2 gene is located on Chr 6p21.33, a disease-susceptibility locus for diffuse panbronchiolitis also encoding mucin21 and mucin-like genes. Specific transcription of the SFTA2 gene in the human lung was verified by RT-PCR. SFTA2 encodes a small hydrophilic precursor peptide (78 amino acids) releasing a 59 amino acid mature peptide after cleavage of an amino-terminal secretory signal. Human SFTA2 has no recognizable homology to other human proteins while orthologues are present in all mammals. Specific antibodies raised against the human peptide recognized a 13 kDa Peptide in human respiratory epithelial cells with a shift to the expected 6 kDa after deglycosylation. In immunofluorescence staining of human lung sections, signals corresponding to SFTA2 were specifically detected in Type II pneumycytes of the alveolar epithelium. While SFTA2 showed no colocalization with lamellar bodies, it was detected in human broncho-alveolar lavage fluid thus establishing this peptide as a novel hydrophilic surfactant protein. In the mouse, Sfta2 was significantly downregulated after induction of a pulmonary inflammatory reaction by intratracheal administration of lipopolysaccharides indicating regulated secretion. We speculate about a possible role of SFTA2 in innate immune defence and inflammation.
    Pediatric Research 11/2011; 70:44-44. DOI:10.1038/pr.2011.269 · 2.31 Impact Factor
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    ABSTRACT: Defects of the NKX2-1 gene, encoding thyroid transcription factor-1, cause brain-thyroid-lung syndrome (MIM 610978), characterised by benign hereditary chorea, congenital hypothyroidism and respiratory disease. The case of a term infant with mild primary congenital hypothyroidism and neonatal persistent respiratory failure with fatal outcome at 10 months of age despite continuous ventilatory support is described. Congenital defects of genes known to disturb surfactant protein and lipid homeostasis (SFTPB, SFTPC, ABCA3) were excluded. Hypothyroidism prompted sequencing of NKX2-1, which revealed a heterozygous 29 bp deletion (c.278_306del29) disrupting the affected allele. Analysis of bronchoalveolar lavage fluid demonstrated an abnormally low amount of surfactant protein C (SP-C) in relation to SP-B, and low levels of surfactant phospholipids, indicating disturbance of SP and lipid homeostasis as a consequence of NKX2-1 haploinsufficiency. NKX2-1 haploinsufficiency may lead to lethal respiratory failure of the newborn due to disruption of pulmonary surfactant homeostasis. NKX2-1 gene analysis should be considered when investigating irreversible respiratory insufficiency of the newborn.
    Archives of Disease in Childhood - Fetal and Neonatal Edition 11/2011; 96(6):F453-6. DOI:10.1136/adc.2009.180448 · 3.12 Impact Factor
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    ABSTRACT: ABCA3 transporter (ATP-binding cassette transporter of the A subfamily) is localized to the limiting membrane of lamellar bodies, organelles for assembly and storage of pulmonary surfactant in alveolar epithelial type II cells (AECII). It transports surfactant phospholipids into lamellar bodies and absence of ABCA3 function disrupts lamellar body biogenesis. Mutations of the ABCA3 gene lead to fatal neonatal surfactant deficiency and chronic interstitial lung disease (ILD) of children. ABCA3 mutations can result in either functional defects of the correctly localized ABCA3 or trafficking/folding defects where mutated ABCA3 remains in the endoplasmic reticulum (ER). Human alveolar epithelial A549 cells were transfected with vectors expressing wild-type ABCA3 or one of the three ABCA3 mutant forms, R43L, R280C and L101P, C-terminally tagged with YFP or hemagglutinin-tag. Localization/trafficking properties were analyzed by immunofluorescence and ABCA3 deglycosylation. Uptake of fluorescent NBD-labeled lipids into lamellar bodies was used as a functional assay. ER stress and apoptotic signaling were examined through RT-PCR based analyses of XBP1 splicing, immunoblotting or FACS analyses of stress/apoptosis proteins, Annexin V surface staining and determination of the intracellular glutathion level. We demonstrate that two ABCA3 mutations, which affect ABCA3 protein trafficking/folding and lead to partial (R280C) or complete (L101P) retention of ABCA3 in the ER compartment, can elevate ER stress and susceptibility to it and induce apoptotic markers in the cultured lung epithelial A549 cells. R43L mutation, resulting in a functional defect of the properly localized ABCA3, had no effect on intracellular stress and apoptotic signaling. Our data suggest that expression of partially or completely ER localized ABCA3 mutant proteins can increase the apoptotic cell death of the affected cells, which are factors that might contribute to the pathogenesis of genetic ILD.
    Respiratory research 01/2011; 12(1):4. DOI:10.1186/1465-9921-12-4 · 3.09 Impact Factor
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    S Herber-Jonat · M Huppmann · R Mittal · M Hammel · A Holzinger · A W Flemmer ·
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    ABSTRACT: Background: Homozygous ABCA3-deficiency is associated with a decreased surfactant function and results in irreversible respiratory failure or in chronic interstitial lung disease. Heterozygous individuals are susceptible for respiratory failure when an additional surfactant-protein-C-deficiency is present.Aim: Determination of lung mechanics and inflammation in heterozygous ABCA3 mice (HZ) after exposure to 95%-oxygen or room-air as compared to wild type mice (WT).Methods: HZ and WT were randomly assigned to 95%-oxygen (n=51) or room air (n=40). One set of mice was ventilated for 25 min and conventional mechanics were measured (Crs, Cstat, Rrs, total lung capacity (TLC), additionally lungimpedance was measured by forced oscillation technique (FOT è Rn, HL, GL, eta). Cell count and interleukins in BAL and wet to dry ratio were determined in ventilated and non-ventilated animals.Results: There was no difference between ventilated HZ and WT mice with regard to respiratory mechanics, impedance measurements and BAL parameters after exposure to room air. After exposure to 95% oxygen TLC was significantly higher in HZ whereas Rn was lower as compared to WT (TLC: 86.6 vs 60.2, p=0.011; Rn: 0.29 vs 0.34, p=0.034). Leucocytes were higher in non-ventilated, room-air housed HZ as compared to WT (310 vs 170/μl, p=0,021). Oxygen exposure followed by ventilation resulted in a significant increase of interleukines HZ (II-6: 3.04 vs 2.55pg/ml, p=0.003; MIP-2: 23.99 vs 11.19, p=0.007).Summary: Our data suggest that lung mechanics and inflammation in HZ are not affected a priori. However, ventilation or oxygen exposure resulted in a phenotypical difference between genotypes.
    Pediatric Research 11/2010; 68:21-22. DOI:10.1203/00006450-201011001-00036 · 2.31 Impact Factor
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    ABSTRACT: Peroxisomes are particularly abundant in the liver and are involved in bile salt synthesis and fatty acid metabolism. Peroxisomal membrane proteins (PMPs) are required for peroxisome biogenesis [e.g., the interacting peroxisomal biogenesis factors Pex13p and Pex14p] and its metabolic function [e.g., the adenosine triphosphate-binding cassette transporters adrenoleukodystrophy protein (ALDP) and PMP70]. Impaired function of PMPs is the underlying cause of Zellweger syndrome and X-linked adrenoleukodystrophy. Here we studied for the first time the putative association of PMPs with cholesterol-enriched lipid rafts and their function in peroxisome biogenesis. Lipid rafts were isolated from Triton X-100-lysed or Lubrol WX-lysed HepG2 cells and analyzed for the presence of various PMPs by western blotting. Lovastatin and methyl-beta-cyclodextrin were used to deplete cholesterol and disrupt lipid rafts in HepG2 cells, and this was followed by immunofluorescence microscopy to determine the subcellular location of catalase and PMPs. Cycloheximide was used to inhibit protein synthesis. Green fluorescent protein-tagged fragments of PMP70 and ALDP were analyzed for their lipid raft association. PMP70 and Pex14p were associated with Triton X-100-resistant rafts, ALDP was associated with Lubrol WX-resistant rafts, and Pex13p was not lipid raft-associated in HepG2 cells. The minimal peroxisomal targeting signals in ALDP and PMP70 were not sufficient for lipid raft association. Cholesterol depletion led to dissociation of PMPs from lipid rafts and impaired sorting of newly synthesized catalase and ALDP but not Pex14p and PMP70. Repletion of cholesterol to these cells efficiently reestablished the peroxisomal sorting of catalase but not ALDP. Conclusion: Human PMPs are differentially associated with lipid rafts independently of the protein homology and/or their functional interaction. Cholesterol is required for peroxisomal lipid raft assembly and peroxisome biogenesis.
    Hepatology 08/2010; 52(2):623-33. DOI:10.1002/hep.23684 · 11.06 Impact Factor
  • L. v. Bomhard · H.-G. Münch · R. A. Mittal · F. Hoffmann · A. Holzinger ·
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    ABSTRACT: Zusammenfassung Hintergrund Milde systemische Hypothermie reduziert Mortalität und neurologische Schäden bei neonataler hypoxisch-ischämischer Enzephalopathie. Wir berichten über unsere Erfahrungen mit Hypothermie als Standardtherapie über 3 Jahre. Patienten und Methoden Von 56 asphyktischen Neugeborenen (Gestationsalter >35 Wochen) wurde bei 33, die innerhalb von 6 h eingetroffen waren, klinische und elektroenzephalographische Zeichen einer Enzephalopathie und keine Kontraindikation aufwiesen, mithilfe einer Kühlmatte eine Körpertemperatur von 33–34°C für 72 h angestrebt. Ergebnisse Insgesamt 29 Neugeborene wurden für 72 h gekühlt, bei 4 Kindern wurde die Behandlung vorzeitig abgebrochen. Die Hypothermiebehandlung ließ sich einfach durchführen. Unerwünschte Ereignisse – vor allem Gerinnungsprobleme, kardiorespiratorische und Nierenfunktionsstörungen – konnten von Folgen der Grunderkrankung nicht abgegrenzt werden. Schlussfolgerungen Die therapeutische Hypothermie scheint eine im klinischen Alltag gut durchführbare Methode zur Neuroprotektion bei hypoxisch-ischämischer Enzephalopathie zu sein. Weitere kontrollierte Studien zur Therapieoptimierung bei Neugeborenen und für eine mögliche Anwendung in anderen Altersgruppen werden benötigt.
    Notfall 08/2010; 13(5):375-380. DOI:10.1007/s10049-010-1319-6 · 0.47 Impact Factor
  • R Mittal · L Jin · M Hammel · G Liebisch · G Schmitz · A Holzinger ·

    Klinische Pädiatrie 06/2010; 222. DOI:10.1055/s-0030-1261416 · 1.06 Impact Factor

  • Klinische Pädiatrie 06/2010; 222:S42-S42. · 1.06 Impact Factor
  • R Polanetz · F Wagner · T Herzinger · A Holzinger ·

    Klinische Pädiatrie 06/2010; 222(S 01). DOI:10.1055/s-0030-1261554 · 1.06 Impact Factor
  • R. Polanetz · F. Wagner · T. Herzinger · A. Holzinger ·

    Klinische Pädiatrie 06/2010; 222:S84-S84. · 1.06 Impact Factor
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    ABSTRACT: Campomelic dysplasia is a malformation syndrome with multiple symptoms including characteristic shortness and bowing of the long bones (campomelia). CD, often lethal due to airway malformations, is caused by heterozygous mutations in SOX9, an SRY-related gene regulating testis and chondrocyte development including expression of many cartilage genes such as type II collagen. Male to female sex reversal occurs in the majority of affected individuals with an XY karyotype. A mild form without campomelia exists, in which sex-reversal may be also absent. We report here two novel SOX9 missense mutations in a male (c.495C>G; p.His165Gln) and a female (c.337A>G; p.Met113Val) within the DNA-binding domain leading to non-lethal acampomelic CD. Functional analyses of mutant proteins demonstrate residual DNA-binding and transactivation of SOX9-regulated genes. Combining our data and reports from the literature we postulate a genotype-phenotype correlation: SOX9 mutations allowing for residual function lead to a mild form of CD in which campomelia and sex reversal may be absent.
    Human Mutation 06/2010; 31(6):E1436-44. DOI:10.1002/humu.21238 · 5.14 Impact Factor
  • A Holzinger · A Staffler · AW Flemmer · T Nicolai · M Wegner · M Hammel ·

    Klinische Pädiatrie 06/2010; 222. DOI:10.1055/s-0030-1261547 · 1.06 Impact Factor
  • A. Holzinger · A. Staffler · A. W. Flemmer · T. Nicolai · M. Wegner · M. Hammel ·

    Klinische Pädiatrie 06/2010; 222:S82-S82. · 1.06 Impact Factor
  • S Herber-Jonat · M Huppmann · R Mittal · M Hammel · A Holzinger · AW Flemmer ·

    Klinische Pädiatrie 06/2010; 222. DOI:10.1055/s-0030-1261415 · 1.06 Impact Factor
  • L. Bomhard · H.-G. Münch · R. A. Mittal · F. Hoffmann · A. Holzinger ·

  • M Hammel · M Klein · T Trips · H Priessmann · T Ankermann · Andreas Holzinger ·
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    ABSTRACT: BACKGROUND: Congenital Central Hypoventilation Syndrome (CCHS, Ondine's curse) is a rare syndrome of dysfunction of the autonomic nervous system characterized by a decreased response to hypercarbia requiring mechanical ventilation in most cases. CCHS is an autosomal-dominant disease associated with tumors of neural crest origin, segmental aganglionosis of the colon, and diffuse autonomic dysregulation symptoms. Most cases of CCHS are caused by de novo heterozygous in-frame expansions within in the PHOX2b gene. PATIENTS AND MAIN RESULTS: Here we report two families in which a PHOX2b defect was inherited from an asymptomatic parent. In family 1 an asymptomatic mother carried a mild mutation (15 bp expansion within the polyalanine repeat) also found in her daughter who was symptomatic immediately after birth but did not require mechanical ventilation. In family 2, two newborn infants with respiratory failure due to insufficient respiratory drive requiring mechanical ventilation were born to asymptomatic parents. A 39 pb expansion within the PHOX2b polyalanine repeat was found in one patient in whom DNA was available, but not in blood leukocytes from any parent. Microsatellite analyses confirmed the identity of the parents, such that a germline mosaicism has to be deduced. CONCLUSIONS: Carriers of mild PHOX2b mutations causing disease in their offspring may be asymptomatic; Modifier genes determining the clinical course may exist. Germline mosaicism may lead to CCHS in children from unaffected parents. Genetic counseling should include these variations.
    Klinische Pädiatrie 10/2009; 221(5):286-9. DOI:10.1055/s-0029-1220941 · 1.06 Impact Factor

  • Chemistry and Physics of Lipids 08/2009; 160. DOI:10.1016/j.chemphyslip.2009.06.138 · 2.42 Impact Factor
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    ABSTRACT: Nerve growth factor (NGF), a neurotrophin, is induced in lung cells by proinflammatory cytokines, and has a role in bronchial hyperreactivity and lung tissue repair. Ventilation induced lung injury, on the other hand, is known to increase the levels of proinflammatory cytokines in the lungs. We investigated whether, and to what extent, various degrees of lung injury induced by short-term ventilation affect NGF levels in the lung tissue of adolescent rabbits. The rabbits were randomized to different modes of ventilation: (1) CON: normal ventilation for 30 min; (2) NVT: normal ventilation for 6 hr; (3) HFQ: ventilation for 6 hr at double frequency, but normal tidal volume (VT); and (4) HVT: 6 hr ventilation at double VT but normal frequency. NGF protein was detected in bronchoalveolar lavage fluid (BALF) and lung tissue in all animals. Ventilation for 6 hr significantly increased NGF levels, in both BALF and lung tissue, in the HFQ and HVT groups as compared to control (P < 0.05). The maximum increase in BALF NGF was seen in the HVT group (P = 0.02 vs. CON and NVT groups, and P = 0.05 vs. HFQ). A parallel increase in interleukin 1-beta (IL1-beta) was observed. Expression of the high-affinity NGF-receptor, tropomyosin-related kinase A (TrkA), was also upregulated in these two groups. Injurious modes of mechanical ventilation upregulate NGF and its receptor TrkA in rabbit lungs, and IL1-beta may be a mediator for this response. We speculate that this increase in NGF level may translate into the development of bronchial hyperreactivity.
    Pediatric Pulmonology 07/2009; 44(7):713-9. DOI:10.1002/ppul.21053 · 2.70 Impact Factor

Publication Stats

711 Citations
78.31 Total Impact Points


  • 2008-2013
    • Technische Universität München
      München, Bavaria, Germany
  • 1999-2012
    • Ludwig-Maximilians-University of Munich
      • • Children in the Department of Surgery, Dr. von Hauner Children's Hospital
      • • Department of Paediatrics
      München, Bavaria, Germany
    • University of Vienna
      • Institute of Neurophysiology
      Wien, Vienna, Austria