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  1. Article: Hypoxic Signalling in Tumour Stroma.

    Laitala, Anu / Erler, Janine T

    Frontiers in oncology

    2018  Volume 8, Page(s) 189

    Abstract: Hypoxia is a common feature in solid tumors and is associated with cancer progression. The main regulators of the hypoxic response are hypoxia-inducible transcription factors (HIFs) that guide the cellular adaptation to hypoxia by gene activation. The ... ...

    Abstract Hypoxia is a common feature in solid tumors and is associated with cancer progression. The main regulators of the hypoxic response are hypoxia-inducible transcription factors (HIFs) that guide the cellular adaptation to hypoxia by gene activation. The actual oxygen sensing is performed by HIF prolyl hydroxylases (PHDs) that under normoxic conditions mark the HIF-α subunit for degradation. Cancer progression is not regulated only by the cancer cells themselves but also by the whole tumor microenvironment, which consists of cellular and extracellular components. Hypoxic conditions also affect the stromal compartment, where stromal cells are in close contact with the cancer cells. The important function of HIF in cancer cells has been shown by many animal models and described in hundreds of reviews, but less in known about PHDs and even less PHDs in stromal cells. Here, we review hypoxic signaling in tumors, mainly in the tumor stroma, with a focus on HIFs and PHDs.
    Language English
    Publishing date 2018-05-29
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2649216-7
    ISSN 2234-943X
    ISSN 2234-943X
    DOI 10.3389/fonc.2018.00189
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Deletion of hypoxia-inducible factor prolyl 4-hydroxylase 2 in FoxD1-lineage mesenchymal cells leads to congenital truncal alopecia.

    Rosendahl, Ann-Helen / Monnius, Mia / Laitala, Anu / Railo, Antti / Miinalainen, Ilkka / Heljasvaara, Ritva / Mäki, Joni M / Myllyharju, Johanna

    The Journal of biological chemistry

    2022  Volume 298, Issue 4, Page(s) 101787

    Abstract: Hypoxia-inducible factors (HIFs) induce numerous genes regulating oxygen homeostasis. As oxygen sensors of the cells, the HIF prolyl 4-hydroxylases (HIF-P4Hs) regulate the stability of HIFs in an oxygen-dependent manner. During hair follicle (HF) ... ...

    Abstract Hypoxia-inducible factors (HIFs) induce numerous genes regulating oxygen homeostasis. As oxygen sensors of the cells, the HIF prolyl 4-hydroxylases (HIF-P4Hs) regulate the stability of HIFs in an oxygen-dependent manner. During hair follicle (HF) morphogenesis and cycling, the location of dermal papilla (DP) alternates between the dermis and hypodermis and results in varying oxygen levels for the DP cells. These cells are known to express hypoxia-inducible genes, but the role of the hypoxia response pathway in HF development and homeostasis has not been studied. Using conditional gene targeting and analysis of hair morphogenesis, we show here that lack of Hif-p4h-2 in Forkhead box D1 (FoxD1)-lineage mesodermal cells interferes with the normal HF development in mice. FoxD1-lineage cells were found to be mainly mesenchymal cells located in the dermis of truncal skin, including those cells composing the DP of HFs. We found that upon Hif-p4h-2 inactivation, HF development was disturbed during the first catagen leading to formation of epithelial-lined HF cysts filled by unorganized keratins, which eventually manifested as truncal alopecia. Furthermore, the depletion of Hif-p4h-2 led to HIF stabilization and dysregulation of multiple genes involved in keratin formation, HF differentiation, and HIF, transforming growth factor β (TGF-β), and Notch signaling. We hypothesize that the failure of HF cycling is likely to be mechanistically caused by disruption of the interplay of the HIF, TGF-β, and Notch pathways. In summary, we show here for the first time that HIF-P4H-2 function in FoxD1-lineage cells is essential for the normal development and homeostasis of HFs.
    MeSH term(s) Alopecia/enzymology ; Alopecia/genetics ; Animals ; Hypoxia-Inducible Factor-Proline Dioxygenases/genetics ; Hypoxia-Inducible Factor-Proline Dioxygenases/metabolism ; Mice ; Oxygen/metabolism ; Transforming Growth Factor beta
    Chemical Substances Foxd1 protein, mouse ; Transforming Growth Factor beta ; Egln1 protein, mouse (EC 1.14.11.29) ; Hypoxia-Inducible Factor-Proline Dioxygenases (EC 1.14.11.29) ; Oxygen (S88TT14065)
    Language English
    Publishing date 2022-03-02
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1016/j.jbc.2022.101787
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  3. Article ; Online: Contribution of HIF-P4H isoenzyme inhibition to metabolism indicates major beneficial effects being conveyed by HIF-P4H-2 antagonism.

    Tapio, Joona / Halmetoja, Riikka / Dimova, Elitsa Y / Mäki, Joni M / Laitala, Anu / Walkinshaw, Gail / Myllyharju, Johanna / Serpi, Raisa / Koivunen, Peppi

    The Journal of biological chemistry

    2022  Volume 298, Issue 8, Page(s) 102222

    Abstract: Hypoxia-inducible factor (HIF) prolyl 4-hydroxylases (HIF-P4Hs 1-3) are druggable targets in renal anemia, where pan-HIF-P4H inhibitors induce an erythropoietic response. Preclinical data suggest that HIF-P4Hs could also be therapeutic targets for ... ...

    Abstract Hypoxia-inducible factor (HIF) prolyl 4-hydroxylases (HIF-P4Hs 1-3) are druggable targets in renal anemia, where pan-HIF-P4H inhibitors induce an erythropoietic response. Preclinical data suggest that HIF-P4Hs could also be therapeutic targets for treating metabolic dysfunction, although the contributions of HIF-P4H isoenzymes in various tissues to the metabolic phenotype are inadequately understood. Here, we used mouse lines that were gene-deficient for HIF-P4Hs 1 to 3 and two preclinical pan-HIF-P4H inhibitors to study the contributions of these isoenzymes to the anthropometric and metabolic outcome and HIF response. We show both inhibitors induced a HIF response in wildtype white adipose tissue (WAT), liver, and skeletal muscle and alleviated metabolic dysfunction during a 6-week treatment period, but they did not alter healthy metabolism. Our data indicate that HIF-P4H-1 contributed especially to skeletal muscle and WAT metabolism and that its loss lowered body weight and serum cholesterol levels upon aging. In addition, we found HIF-P4H-3 had effects on the liver and WAT and its loss increased body weight, adiposity, liver weight and triglyceride levels, WAT inflammation, and cholesterol levels and resulted in hyperglycemia and insulin resistance, especially during aging. Finally, we demonstrate HIF-P4H-2 affected all tissues studied; its inhibition lowered body and liver weight and serum cholesterol levels and improved glucose tolerance. We found very few HIF target metabolic mRNAs were regulated by the inhibition of three isoenzymes, thus suggesting a potential for selective therapeutic tractability. Altogether, these data provide specifications for the future development of HIF-P4H inhibitors for the treatment of metabolic diseases.
    MeSH term(s) Adipose Tissue, White/metabolism ; Aging/metabolism ; Animals ; Body Weight ; Cholesterol/blood ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism ; Hypoxia-Inducible Factor-Proline Dioxygenases/antagonists & inhibitors ; Hypoxia-Inducible Factor-Proline Dioxygenases/metabolism ; Insulin Resistance ; Isoenzymes/antagonists & inhibitors ; Isoenzymes/metabolism ; Liver/metabolism ; Mice ; Muscle, Skeletal/metabolism ; Obesity/metabolism
    Chemical Substances Hypoxia-Inducible Factor 1, alpha Subunit ; Isoenzymes ; Cholesterol (97C5T2UQ7J) ; Hypoxia-Inducible Factor-Proline Dioxygenases (EC 1.14.11.29)
    Language English
    Publishing date 2022-07-01
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1016/j.jbc.2022.102222
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  4. Article ; Online: Suppression of tumor-associated neutrophils by lorlatinib attenuates pancreatic cancer growth and improves treatment with immune checkpoint blockade.

    Nielsen, Sebastian R / Strøbech, Jan E / Horton, Edward R / Jackstadt, Rene / Laitala, Anu / Bravo, Marina C / Maltese, Giorgia / Jensen, Adina R D / Reuten, Raphael / Rafaeva, Maria / Karim, Saadia A / Hwang, Chang-Il / Arnes, Luis / Tuveson, David A / Sansom, Owen J / Morton, Jennifer P / Erler, Janine T

    Nature communications

    2021  Volume 12, Issue 1, Page(s) 3414

    Abstract: Pancreatic ductal adenocarcinoma (PDAC) patients have a 5-year survival rate of only 8% largely due to late diagnosis and insufficient therapeutic options. Neutrophils are among the most abundant immune cell type within the PDAC tumor microenvironment ( ... ...

    Abstract Pancreatic ductal adenocarcinoma (PDAC) patients have a 5-year survival rate of only 8% largely due to late diagnosis and insufficient therapeutic options. Neutrophils are among the most abundant immune cell type within the PDAC tumor microenvironment (TME), and are associated with a poor clinical prognosis. However, despite recent advances in understanding neutrophil biology in cancer, therapies targeting tumor-associated neutrophils are lacking. Here, we demonstrate, using pre-clinical mouse models of PDAC, that lorlatinib attenuates PDAC progression by suppressing neutrophil development and mobilization, and by modulating tumor-promoting neutrophil functions within the TME. When combined, lorlatinib also improves the response to anti-PD-1 blockade resulting in more activated CD8 + T cells in PDAC tumors. In summary, this study identifies an effect of lorlatinib in modulating tumor-associated neutrophils, and demonstrates the potential of lorlatinib to treat PDAC.
    MeSH term(s) Aminopyridines ; Animals ; Antineoplastic Combined Chemotherapy Protocols/pharmacology ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use ; CD8-Positive T-Lymphocytes/drug effects ; CD8-Positive T-Lymphocytes/immunology ; Carcinoma, Pancreatic Ductal/drug therapy ; Carcinoma, Pancreatic Ductal/pathology ; Cell Line, Tumor/transplantation ; Disease Models, Animal ; Drug Synergism ; Female ; Humans ; Immune Checkpoint Inhibitors/pharmacology ; Immune Checkpoint Inhibitors/therapeutic use ; Lactams ; Lactams, Macrocyclic/pharmacology ; Lactams, Macrocyclic/therapeutic use ; Lymphocyte Activation/drug effects ; Lymphocytes, Tumor-Infiltrating/drug effects ; Lymphocytes, Tumor-Infiltrating/immunology ; Male ; Mice ; Mice, Transgenic ; Neutrophils/drug effects ; Neutrophils/immunology ; Pancreatic Neoplasms/drug therapy ; Pancreatic Neoplasms/genetics ; Pancreatic Neoplasms/immunology ; Pancreatic Neoplasms/pathology ; Programmed Cell Death 1 Receptor/antagonists & inhibitors ; Programmed Cell Death 1 Receptor/metabolism ; Pyrazoles ; Tumor Microenvironment/drug effects ; Tumor Microenvironment/immunology
    Chemical Substances Aminopyridines ; Immune Checkpoint Inhibitors ; Lactams ; Lactams, Macrocyclic ; Pdcd1 protein, mouse ; Programmed Cell Death 1 Receptor ; Pyrazoles ; lorlatinib (OSP71S83EU)
    Language English
    Publishing date 2021-06-07
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2553671-0
    ISSN 2041-1723 ; 2041-1723
    ISSN (online) 2041-1723
    ISSN 2041-1723
    DOI 10.1038/s41467-021-23731-7
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  5. Article ; Online: Basement membrane stiffness determines metastases formation.

    Reuten, Raphael / Zendehroud, Sina / Nicolau, Monica / Fleischhauer, Lutz / Laitala, Anu / Kiderlen, Stefanie / Nikodemus, Denise / Wullkopf, Lena / Nielsen, Sebastian Rune / McNeilly, Sarah / Prein, Carina / Rafaeva, Maria / Schoof, Erwin M / Furtwängler, Benjamin / Porse, Bo T / Kim, Hyobin / Won, Kyoung Jae / Sudhop, Stefanie / Zornhagen, Kamilla Westarp /
    Suhr, Frank / Maniati, Eleni / Pearce, Oliver M T / Koch, Manuel / Oddershede, Lene Broeng / Van Agtmael, Tom / Madsen, Chris D / Mayorca-Guiliani, Alejandro E / Bloch, Wilhelm / Netz, Roland R / Clausen-Schaumann, Hauke / Erler, Janine T

    Nature materials

    2021  Volume 20, Issue 6, Page(s) 892–903

    Abstract: The basement membrane (BM) is a special type of extracellular matrix and presents the major barrier cancer cells have to overcome multiple times to form metastases. Here we show that BM stiffness is a major determinant of metastases formation in several ... ...

    Abstract The basement membrane (BM) is a special type of extracellular matrix and presents the major barrier cancer cells have to overcome multiple times to form metastases. Here we show that BM stiffness is a major determinant of metastases formation in several tissues and identify netrin-4 (Net4) as a key regulator of BM stiffness. Mechanistically, our biophysical and functional analyses in combination with mathematical simulations show that Net4 softens the mechanical properties of native BMs by opening laminin node complexes, decreasing cancer cell potential to transmigrate this barrier despite creating bigger pores. Our results therefore reveal that BM stiffness is dominant over pore size, and that the mechanical properties of 'normal' BMs determine metastases formation and patient survival independent of cancer-mediated alterations. Thus, identifying individual Net4 protein levels within native BMs in major metastatic organs may have the potential to define patient survival even before tumour formation. The ratio of Net4 to laminin molecules determines BM stiffness, such that the more Net4, the softer the BM, thereby decreasing cancer cell invasion activity.
    MeSH term(s) Basement Membrane/metabolism ; Biomechanical Phenomena ; Cell Line, Tumor ; Humans ; Mechanical Phenomena ; Neoplasm Metastasis ; Netrins/metabolism
    Chemical Substances Netrins
    Language English
    Publishing date 2021-01-25
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2088679-2
    ISSN 1476-4660 ; 1476-1122
    ISSN (online) 1476-4660
    ISSN 1476-1122
    DOI 10.1038/s41563-020-00894-0
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  6. Article ; Online: Muscle composition is regulated by a Lox-TGFβ feedback loop.

    Kutchuk, Liora / Laitala, Anu / Soueid-Bomgarten, Sharon / Shentzer, Pessia / Rosendahl, Ann-Helen / Eilot, Shelly / Grossman, Moran / Sagi, Irit / Sormunen, Raija / Myllyharju, Johanna / Mäki, Joni M / Hasson, Peleg

    Development (Cambridge, England)

    2015  Volume 142, Issue 5, Page(s) 983–993

    Abstract: Muscle is an integrated tissue composed of distinct cell types and extracellular matrix. While much emphasis has been placed on the factors required for the specification of the cells that comprise muscle, little is known about the crosstalk between them ...

    Abstract Muscle is an integrated tissue composed of distinct cell types and extracellular matrix. While much emphasis has been placed on the factors required for the specification of the cells that comprise muscle, little is known about the crosstalk between them that enables the development of a patterned and functional tissue. We find in mice that deletion of lysyl oxidase (Lox), an extracellular enzyme regulating collagen maturation and organization, uncouples the balance between the amount of myofibers and that of muscle connective tissue (MCT). We show that Lox secreted from the myofibers attenuates TGFβ signaling, an inhibitor of myofiber differentiation and promoter of MCT development. We further demonstrate that a TGFβ-Lox feedback loop between the MCT and myofibers maintains the dynamic developmental homeostasis between muscle components while also regulating MCT organization. Our results allow a better understanding of diseases such as Duchenne muscular dystrophy, in which LOX and TGFβ signaling have been implicated and the balance between muscle constituents is disturbed.
    MeSH term(s) Animals ; Connective Tissue/embryology ; Connective Tissue/metabolism ; Connective Tissue/ultrastructure ; Extracellular Matrix Proteins/genetics ; Extracellular Matrix Proteins/metabolism ; Female ; Immunohistochemistry ; In Situ Hybridization ; Mice ; Microscopy, Electron, Transmission ; Muscles/embryology ; Muscles/metabolism ; Muscles/ultrastructure ; Pregnancy ; Protein-Lysine 6-Oxidase/genetics ; Protein-Lysine 6-Oxidase/metabolism ; Signal Transduction/genetics ; Signal Transduction/physiology ; Transforming Growth Factor beta/genetics ; Transforming Growth Factor beta/metabolism
    Chemical Substances Extracellular Matrix Proteins ; Transforming Growth Factor beta ; Lox protein, mouse (149137-54-2) ; Protein-Lysine 6-Oxidase (EC 1.4.3.13)
    Language English
    Publishing date 2015-03-01
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 90607-4
    ISSN 1477-9129 ; 0950-1991
    ISSN (online) 1477-9129
    ISSN 0950-1991
    DOI 10.1242/dev.113449
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  7. Article ; Online: Transmembrane prolyl 4-hydroxylase is a fourth prolyl 4-hydroxylase regulating EPO production and erythropoiesis.

    Laitala, Anu / Aro, Ellinoora / Walkinshaw, Gail / Mäki, Joni M / Rossi, Maarit / Heikkilä, Minna / Savolainen, Eeva-Riitta / Arend, Michael / Kivirikko, Kari I / Koivunen, Peppi / Myllyharju, Johanna

    Blood

    2012  Volume 120, Issue 16, Page(s) 3336–3344

    Abstract: An endoplasmic reticulum transmembrane prolyl 4-hydroxylase (P4H-TM) is able to hydroxylate the α subunit of the hypoxia-inducible factor (HIF) in vitro and in cultured cells, but nothing is known about its roles in mammalian erythropoiesis. We studied ... ...

    Abstract An endoplasmic reticulum transmembrane prolyl 4-hydroxylase (P4H-TM) is able to hydroxylate the α subunit of the hypoxia-inducible factor (HIF) in vitro and in cultured cells, but nothing is known about its roles in mammalian erythropoiesis. We studied such roles here by administering a HIF-P4H inhibitor, FG-4497, to P4h-tm(-/-) mice. This caused larger increases in serum Epo concentration and kidney but not liver Hif-1α and Hif-2α protein and Epo mRNA levels than in wild-type mice, while the liver Hepcidin mRNA level was lower in the P4h-tm(-/-) mice than in the wild-type. Similar, but not identical, differences were also seen between FG-4497-treated Hif-p4h-2 hypomorphic (Hif-p4h-2(gt/gt)) and Hif-p4h-3(-/-) mice versus wild-type mice. FG-4497 administration increased hemoglobin and hematocrit values similarly in the P4h-tm(-/-) and wild-type mice, but caused higher increases in both values in the Hif-p4h-2(gt/gt) mice and in hematocrit value in the Hif-p4h-3(-/-) mice than in the wild-type. Hif-p4h-2(gt/gt)/P4h-tm(-/-) double gene-modified mice nevertheless had increased hemoglobin and hematocrit values without any FG-4497 administration, although no such abnormalities were seen in the Hif-p4h-2(gt/gt) or P4h-tm(-/-) mice. Our data thus indicate that P4H-TM plays a role in the regulation of EPO production, hepcidin expression, and erythropoiesis.
    MeSH term(s) Animals ; Antimicrobial Cationic Peptides/metabolism ; Blotting, Western ; Enzyme Inhibitors/pharmacology ; Enzyme-Linked Immunosorbent Assay ; Erythropoiesis/physiology ; Erythropoietin/blood ; Female ; Hematocrit ; Hemoglobins/metabolism ; Hepcidins ; Hypoxia-Inducible Factor 1, alpha Subunit/antagonists & inhibitors ; Hypoxia-Inducible Factor 1, alpha Subunit/physiology ; Liver/metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Procollagen-Proline Dioxygenase/antagonists & inhibitors ; Procollagen-Proline Dioxygenase/physiology ; RNA, Messenger/genetics ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction
    Chemical Substances Antimicrobial Cationic Peptides ; Enzyme Inhibitors ; Hamp protein, mouse ; Hemoglobins ; Hepcidins ; Hif1a protein, mouse ; Hypoxia-Inducible Factor 1, alpha Subunit ; RNA, Messenger ; Erythropoietin (11096-26-7) ; Procollagen-Proline Dioxygenase (EC 1.14.11.2)
    Language English
    Publishing date 2012-09-05
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80069-7
    ISSN 1528-0020 ; 0006-4971
    ISSN (online) 1528-0020
    ISSN 0006-4971
    DOI 10.1182/blood-2012-07-441824
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