Change search
Link to record
Permanent link

Direct link
Holback, Sofia
Publications (7 of 7) Show all publications
Holback, S. (2009). Proteolytic processing of the Alzheimer APP protein family during neuronal differentiation. (Doctoral dissertation). Stockholm: Department of Neurochemistry, Stockholm University
Open this publication in new window or tab >>Proteolytic processing of the Alzheimer APP protein family during neuronal differentiation
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Increased amyloid-β (Aβ) load in the brain, neurite degeneration, neuronal loss, and decreased levels of several neurotrophins are among the characteristics of Alzheimer’s disease (AD). Generation of Aβ occurs when the amyloid precursor protein (APP) is proteolytically processed by β- and γ-secretases in the amyloidogenic pathway. However, Aβ formation is prevented if APP is cleaved by α- and γ- secretases in the non-amyloidogenic pathway. The normal function of APP is still not fully known. It seems clear that the different fragments that are produced during proteolytic processing have different bioactive properties. APP and its metabolites have been implicated in neurite outgrowth, synaptogenesis, cell adhesion, neuroprotection and apoptosis.

The aim of this thesis was to investigate how neurotrophic factors affect the synthesis and processing of APP and its two mammalian paralogues the APP-like protein-1 and-2 (APLP1 and APLP2). We also wanted to determine how the expression levels of α- and β- secretases were affected in response to these factors. In addition, we wanted to analyze if the levels and function of the most well characterized APP adaptor protein, Fe65, was regulated during neuronal differentiation.

Our results show that retinoic acid (RA), insulin-like growth factor-1 (IGF-1), and brain derived neurotrophic factor (BDNF) all regulate expression levels and processing of the APP protein family. Interestingly, the increased processing of the APP family involves different signaling pathways. The PI3-K/Akt pathway is involved in IGF-1-induced APP and APLP1, but not APLP2, processing. In addition, RA-induced expression of the α-secretase, a disintegrin and metalloproteinase (ADAM) 10 is dependent on PI3-K, whereas PKC is involved in RA-induced expression of another α-secretase, ADAM17/TACE. Furthermore, we present evidence that maturation of the adaptor protein Fe65, as well as its docking to APP, increases concomitant with neuronal differentiation.

Place, publisher, year, edition, pages
Stockholm: Department of Neurochemistry, Stockholm University, 2009. p. 82
Keywords
APP, APLP1, APLP2, differentiation, processing, retinoic acid, secretase
National Category
Neurosciences
Research subject
Neurochemistry and Molecular Neurobiology
Identifiers
urn:nbn:se:su:diva-31301 (URN)978-91-7155-942-5 (ISBN)
Public defence
2009-12-18, Magnélisalen, Kemiska övningslaboratioriet, Svante Arrhenius väg 16 B, Stockholm, 10:00 (English)
Opponent
Supervisors
Note
At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 4: Manuscript.Available from: 2009-11-26 Created: 2009-11-10 Last updated: 2022-02-25Bibliographically approved
Holback, S., Adlerz, L., Gatsinzi, T., Jacobsen, K. T. & Iverfeldt, K. (2008). PI3-K- and PKC-dependent up-regulation of APP processing enzymes by retinoic acid. Biochemical and Biophysical Research Communications - BBRC, 365(2), 298-303
Open this publication in new window or tab >>PI3-K- and PKC-dependent up-regulation of APP processing enzymes by retinoic acid
Show others...
2008 (English)In: Biochemical and Biophysical Research Communications - BBRC, ISSN 0006-291X, E-ISSN 1090-2104, Vol. 365, no 2, p. 298-303Article in journal (Refereed) Published
Abstract [en]

Retinoic acid stimulates α-secretase processing of amyloid precursor protein (APP) and decreases β-secretase cleavage that leads to amyloid-β formation. Here, we investigated the effect of retinoic acid on the two putative α-secretases, the disintegrin metalloproteinases ADAM10 and TACE, and the β-site cleaving enzyme BACE1, in human neuroblastoma SH-SY5Y cells. Western blot analysis showed that exposure to retinoic acid resulted in significantly increased levels of ADAM10 and TACE, suggesting that regulation of α-secretases causes the effects on APP processing. The presence of the phosphatidylinositol 3-kinase inhibitor LY 294002 selectively reduced the effect on ADAM10 protein levels but not on ADAM10 mRNA levels as determined by RT-PCR. On the other hand, the effect on TACE was shown to be dependent on protein kinase C, since it was completely blocked in the presence of the inhibitor bisindolylmaleimide XI. Our data indicate that different signalling pathways are involved in retinoic acid-induced up-regulation of the secretases.

Keywords
ADAM10, APP, BACE1, BDNF, PI3-kinase, Protein kinase C, Retinoic acid, TACE
National Category
Natural Sciences
Identifiers
urn:nbn:se:su:diva-19985 (URN)10.1016/j.bbrc.2007.10.167 (DOI)000251494000015 ()
Available from: 2009-01-23 Created: 2009-01-23 Last updated: 2022-02-25Bibliographically approved
Adlerz, L., Holback, S., Multhaup, G. & Iverfeldt, K. (2007). IGF-1-induced Processing of the Amyloid Precursor Protein Family Is Mediated by Different Signaling Pathways. Journal of Biological Chemistry, 282(14), 10203-10209
Open this publication in new window or tab >>IGF-1-induced Processing of the Amyloid Precursor Protein Family Is Mediated by Different Signaling Pathways
2007 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 282, no 14, p. 10203-10209Article in journal (Refereed) Published
Abstract [en]

The mammalian amyloid precursor protein (APP) protein family consists of the APP and the amyloid precursor-like proteins 1 and 2 (APLP1 and APLP2). The neurotoxic amyloid beta-peptide (Abeta) originates from APP, which is the only member of this protein family implicated in Alzheimer disease. However, the three homologous proteins have been proposed to be processed in similar ways and to have essential and overlapping functions. Therefore, it is also important to take into account the effects on the processing and function of the APP-like proteins in the development of therapeutic drugs aimed at decreasing the production of Abeta. Insulin and insulin-like growth factor-1 (IGF-1) have been shown to regulate APP processing and the levels of Abeta in the brain. In the present study, we show that IGF-1 increases alpha-secretase processing of endogenous APP and also increases ectodomain shedding of APLP1 and APLP2 in human SH-SY5Y neuroblastoma cells. We also investigated the role of different IGF-1-induced signaling pathways, using specific inhibitors for phosphatidylinositol 3-kinase and mitogen-activated protein kinase (MAPK). Our results indicate that phosphatidylinositol 3-kinase is involved in ectodomain shedding of APP and APLP1, but not APLP2, and that MAPK is involved only in the ectodomain shedding of APLP1.

National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:su:diva-21101 (URN)10.1074/jbc.M611183200 (DOI)000245941000010 ()
Available from: 2007-04-20 Created: 2007-04-20 Last updated: 2022-02-25Bibliographically approved
Axelsson, V., Holback, S., Sjögren, M., Gustafsson, H. & Forsby, A. (2006). Gliotoxin induces caspase-dependent neurite degeneration and calpain-mediated general cytotoxicity in differentiated human neuroblastoma SH-SY5Y cells.. Biochem Biophys Res Commun, 345(3), 1068-74
Open this publication in new window or tab >>Gliotoxin induces caspase-dependent neurite degeneration and calpain-mediated general cytotoxicity in differentiated human neuroblastoma SH-SY5Y cells.
Show others...
2006 (English)In: Biochem Biophys Res Commun, ISSN 0006-291X, Vol. 345, no 3, p. 1068-74Article in journal (Refereed) Published
Abstract [en]

In this study, a significant increase by 50% in intracellular free calcium concentration ([Ca(2+)](i)) was observed in differentiated human neuroblastoma (SH-SY5Y) cells after exposure to 0.25microM of the fungal metabolite gliotoxin for 72h. Further, the involvement of caspases and calpains was demonstrated to underlie the gliotoxin-induced cytotoxic and neurite degenerative effects. The caspase inhibitor Z-VAD-fmk almost completely reduced the neurite degeneration from 40% degeneration of neurites to 5% as compared to control. Inhibition of calpains with calpeptin significantly attenuated gliotoxin-induced cytotoxicity, determined as reduction in total cellular protein content, from 43% to 14% as compared to control cells. Western blot analyses of alphaII-spectrin breakdown fragments confirmed activity of the proteases, and that alphaII-spectrin was cleaved by caspases in gliotoxin-exposed cells. These results show that calpains and caspases have a role in the toxicity of gliotoxin in differentiated SH-SY5Y cells and that the process may be Ca(2+)-mediated.

Keywords
Calcium/metabolism, Calpain/*metabolism, Caspases/*metabolism, Cell Differentiation, Cell Line;/pharmacology, Gliotoxin/*pharmacology, Humans, Immunosuppressive Agents/*pharmacology, Neurites/*metabolism, Neuroblastoma/*metabolism, Spectrin/metabolism
National Category
Pharmacology and Toxicology
Identifiers
urn:nbn:se:su:diva-20495 (URN)16712786 (PubMedID)
Available from: 2007-11-26 Created: 2007-11-26 Last updated: 2022-02-25Bibliographically approved
Holback, S., Adlerz, L. & Iverfeldt, K. (2005). Increased processing of APLP2 and APP with concomitantformation of APP intracellular domains in BDNF and retinoic acid-differentiated human neuroblastoma cells. Journal of Neurochemistry, 95(4), 1059-1068
Open this publication in new window or tab >>Increased processing of APLP2 and APP with concomitantformation of APP intracellular domains in BDNF and retinoic acid-differentiated human neuroblastoma cells
2005 (English)In: Journal of Neurochemistry, ISSN 0022-3042, E-ISSN 1471-4159, Vol. 95, no 4, p. 1059-1068Article in journal (Refereed) Published
Abstract [en]

The amyloid precursor protein (APP) belongs to a conserved gene family, also including the amyloid precursor-like proteins, APLP1 and APLP2. We have previously shown that all members of the APP protein family are up-regulated upon retinoic acid (RA)-induced neuronal differentiation of SH-SY5Y neuroblastoma cells. Here, we demonstrate that RA also affects the processing of APLP2 and APP, as shown by increased shedding of both sAPLP2 and sAPPalpha as well as elevated levels of the APP intracellular domains (AICDs). Brain-derived neurotrophic factor (BDNF) has been reported to induce APP promoter activity and RA induces expression of the tyrosine kinase receptor B (TrkB) in neuroblastoma cells. We show that the increase in shedding of both APLP2 and APP in response to RA is not mediated through the TrkB receptor. However, BDNF concomitant with RA increased the expression of APP even further. In addition, the secretion of sAPLP2 and sAPPalpha, as well as the levels of AICDs increased in response to BDNF. In contrast, the levels of membrane-bound APP C-terminal fragment C99 significantly decreased. Our results suggest that RA and BDNF shifts APP processing towards the alpha-secretase pathway. In addition, we show that RA and BDNF regulate N-linked glycosylation of APLP1.

Keywords
Alzheimer’s disease, amyloid precursor protein, amyloid precursor-like proteins 1 and 2, amyloid precursor protein intracellular domain, processing.
National Category
Biochemistry and Molecular Biology Neurosciences
Identifiers
urn:nbn:se:su:diva-31749 (URN)10.1111/j.1471-4159.2005.03440.x (DOI)000232850000014 ()
Available from: 2009-12-14 Created: 2009-11-26 Last updated: 2022-02-25Bibliographically approved
Adlerz, L., Beckman, M., Holback, S., Tehranian, R., Cortés Toro, V. & Iverfeldt, K. (2003). Accumulation of the amyloid precursor-like protein APLP2 and reduction of APLP1 in retinoic acid-differentiated human neuroblastoma cells upon curcumin-induced neurite retraction. Brain Research. Molecular Brain Research, 119(1), 62-72
Open this publication in new window or tab >>Accumulation of the amyloid precursor-like protein APLP2 and reduction of APLP1 in retinoic acid-differentiated human neuroblastoma cells upon curcumin-induced neurite retraction
Show others...
2003 (English)In: Brain Research. Molecular Brain Research, ISSN 0169-328X, E-ISSN 1872-6941, Vol. 119, no 1, p. 62-72Article in journal (Refereed) Published
Abstract [en]

Amyloid precursor protein (APP) belongs to a conserved gene family, also including the amyloid precursor-like proteins, APLP1 and APLP2. The function of these three proteins is not yet fully understood. One of the proposed roles of APP is to promote neurite outgrowth. The aim of this study was to investigate the regulation of the expression levels of APP family members during neurite outgrowth. We observed that retinoic acid (RA)-induced neuronal differentiation of human SH-SY5Y cells resulted in increased expression of APP, APLP1 and APLP2. We also examined the effect of the NFκB, AP-1 and c-Jun N-terminal kinase inhibitor curcumin (diferuloylmethane) on the RA-induced expression levels of these proteins. We found that treatment with curcumin counteracted the RA-induced mRNA expression of all APP family members. In addition, we observed that curcumin treatment resulted in neurite retraction without any effect on cell viability. Surprisingly, curcumin had differential effects on the APLP protein levels in RA-differentiated cells. RA-induced APLP1 protein expression was blocked by curcumin, while the APLP2 protein levels were further increased. APP protein levels were not affected by curcumin treatment. We propose that the sustained levels of APP and the elevated levels of APLP2, in spite of the reduced mRNA expression, are due to altered proteolytic processing of these proteins. Furthermore, our results suggest that APLP1 does not undergo the same type of regulated processing as APP and APLP2.

Keywords
APP, APLP1, APLP2; Curcumin, Retinoic acid, Neurite outgrowth
National Category
Neurosciences Neurology
Identifiers
urn:nbn:se:su:diva-31748 (URN)10.1016/j.molbrainres.2003.08.014 (DOI)000186670700007 ()
Available from: 2009-12-14 Created: 2009-11-26 Last updated: 2022-02-25Bibliographically approved
Holback, S., Koistinen, N., Jacobsen, K. & Iverfeldt, K.Retinoic acid stimulates maturation of the adaptor protein Fe65 and its binding to the amyloid precursor protein.
Open this publication in new window or tab >>Retinoic acid stimulates maturation of the adaptor protein Fe65 and its binding to the amyloid precursor protein
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Retinoic acid (RA) stimulates both synthesis and processing of the amyloid precursor protein (APP) and its mammalian paralogues, the APP-like proteins 1 and 2 (APLP1 and APLP2). Previously, we have detected increased levels of the APP and APLP1 intracellular dolmans, AICD and ALID1 respectively, concomitant with RA-induced neuronal differentiation. Here we used Western blot analysis to show increased levels of the mature form of the adaptor protein Fe65 during RA- as well as nerve growth factor-induced differentiation. Co-immunoprecipitation studies also revealed that increased binding of Fe65 to APP and APLP1 occurred during neuronal differentiation. Furthermore, exposure to RA decreased the phosphorylation of Thr668 located in the cytoplasmic domain of APP.

Keywords
Fe65, retinoic acid, amyloid precursor protein, neuronal differentiation
National Category
Biochemistry and Molecular Biology
Research subject
Neurochemistry with Molecular Neurobiology
Identifiers
urn:nbn:se:su:diva-32579 (URN)
Available from: 2009-12-14 Created: 2009-12-14 Last updated: 2022-02-25Bibliographically approved
Organisations

Search in DiVA

Show all publications