Cell communication systems based on polypeptide ligands use transmembrane receptors to transmit signals across the plasma membrane. In their biogenesis, receptors depend on the endoplasmic reticulum (ER)-Golgi system for folding, maturation, transport and localization to the cell surface. ER stress, caused by protein overproduction and misfolding, is a well-known pathology in neurodegeneration, cancer and numerous other diseases. How ER stress affects cell communication via transmembrane receptors is largely unknown. In disease models of multiple myeloma, chronic lymphocytic leukemia and osteogenesis imperfecta, we show that ER stress leads to loss of the mature transmembrane receptors FGFR3, ROR1, FGFR1, LRP6, FZD5 and PTH1R at the cell surface, resulting in impaired downstream signaling. This is caused by downregulation of receptor production and increased intracellular retention of immature receptor forms. Reduction of ER stress by treatment of cells with the chemical chaperone tauroursodeoxycholic acid or by expression of the chaperone protein BiP resulted in restoration of receptor maturation and signaling. We show a previously unappreciated pathological effect of ER stress; impaired cellular communication due to altered receptor processing. Our findings have implications for disease mechanisms related to ER stress and are particularly important when receptor-based pharmacological approaches are used for treatment.

Department of Biology Faculty of Medicine Masaryk University 62500 Brno Czech Republic

Department of Cytokinetics Institute of Biophysics Czech Academy of Sciences 61265 Brno Czech Republic

Department of Experimental Biology Faculty of Science Masaryk University 62500 Brno Czech Republic

Department of Histology and Embryology Faculty of Medicine Masaryk University 62500 Brno Czech Republic

Department of Internal Medicine Hematology and Oncology University Hospital Brno 62500 Brno Czech Republic

Department of Orthopaedic Surgery Human Genetics and Obstetrics and Gynecology University of California at Los Angeles Los Angeles CA 90095 USA

Institute of Animal Physiology and Genetics Czech Academy of Sciences 60200 Brno Czech Republic

International Clinical Research Center St Anne's University Hospital 65691 Brno Czech Republic

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Hetz C, Zhang K, Kaufman RJ. Mechanisms, regulation and functions of the unfolded protein response. Nat Rev Mol Cell Biol. 2020;21(8):421–38. PubMed PMC

Wang M, Kaufman RJ. The impact of the endoplasmic reticulum protein-folding environment on cancer development. Nat Rev Cancer. 2014;14(9):581–97. PubMed

Cox JS, Shamu CE, Walter P. Transcriptional induction of genes encoding endoplasmic reticulum resident proteins requires a transmembrane protein kinase. Cell. 1993;73:1197–206. PubMed

Harding HP, Zhang Y, Ron D. Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature. 1999;397:271–4. PubMed

Haze K, Yoshida H, Yanagi H, Yura T, Mori K. Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress. Mol Biol Cell. 1999;10:3787–99. PubMed PMC

Frantz C, Stewart KM, Weaver VM. The extracellular matrix at a glance. J Cell Sci. 2010;123(24):4195–200. PubMed PMC

Moscatelli D. High and low affinity binding sites for basic fibroblast growth factor on cultured cells: absence of a role for low affinity binding in the stimulation of plasminogen activator production by bovine capillary endothelial cells. J Cell Physiol. 1987;131:123–30. PubMed

Liuzzo J, Moscatelli D. Human Leukemia Cell Lines Bind Basic Fibroblast Growth Factor (FGF) on FGF Receptors and Heparan Sulfates: Downmodulation of FGF Receptors by Phorbol Ester. Blood. 1996;87:245–55. PubMed

Blair HC, Larrouture QC, Li Y, Lin H, Beer-Stoltz D, Liu L, et al. Osteoblast Differentiation and Bone Matrix Formation In Vivo and In Vitro. Tissue Eng Part B Rev. 2017;23:268–80. PubMed PMC

Li Y, Kang K, Krahn JM, Croutwater N, Lee K, Umbach DM, et al. A comprehensive genomic pan-cancer classification using The Cancer Genome Atlas gene expression data. BMC Genomics. 2017;3;18(1):508 PubMed PMC

Dai C, Dai S, Cao J. Proteotoxic stress of cancer: implication of the heat-shock response in oncogenesis. J Cell Physiol. 2012;227:2982–7. PubMed PMC

Shi H, O’Reilly VC, Moreau JLM, Bewes TR, Yam MX, Chapman BE, et al. Gestational stress induces the unfolded protein response, resulting in heart defects. Development (Cambridge). 2016;143:2561–72. PubMed

Thangaraj A, Sil S, Tripathi A, Chivero ET, Periyasamy P, Buch S. Targeting endoplasmic reticulum stress and autophagy as therapeutic approaches for neurological diseases. Int Rev Cell Mol Biol. 2020;350:285–325. PubMed

Sprenkle NT, Sims SG, Sánchez CL, Meares GP. Endoplasmic reticulum stress and inflammation in the central nervous system. Mol Neurodegener. 2017;12(1):42. PubMed PMC

Yadav RK, Chae S-W, Kim H-R, Chae HJ. Endoplasmic Reticulum Stress and Cancer. J Cancer Prev. 2014;19:75–88. PubMed PMC

Back SH, Kaufman RJ. Endoplasmic reticulum stress and type 2 diabetes. Annu Rev Biochem. 2012;81:767–93. PubMed PMC

Todd DJ, Lee AH, Glimcher LH. The endoplasmic reticulum stress response in immunity and autoimmunity. Nat Rev Immunol. 2008;8(9):663–74. PubMed

Kazandjian D. Multiple myeloma epidemiology and survival: A unique malignancy. Semin Oncol. 2016;43:676–81. PubMed PMC

Nikesitch N, Lee JM, Ling S, Roberts TL. Endoplasmic reticulum stress in the development of multiple myeloma and drug resistance. Clin Transl Immunology. 2018;7(1):e1007. 10.1002/cti2.1007. PubMed PMC

Obeng EA, Carlson LM, Gutman DM, Harrington WJ, Lee KP, Boise LH. Proteasome inhibitors induce a terminal unfolded protein response in multiple myeloma cells. Blood. 2006;107:4907–16. PubMed PMC

Curran MP, McKeage K. Bortezomib: A review of its use in patients with multiple myeloma. Drugs. 2009;69:859–88. PubMed

Chesi M, Nardini E, Brents LA, Schröck E, Ried T, Kuehl WM, et al. Frequent translocation t(4;14)(p16.3;q32.3) in multiple myeloma is associated with increased expression and activating mutations of fibroblast growth factor receptor 3. Nat Genet. 1997;16:260–4 PubMed PMC

Krejci P, Krakow D, Mekikian PB, Wilcox WR. Fibroblast Growth Factors 1, 2, 17, and 19 Are the Predominant FGF Ligands Expressed in Human Fetal Growth Plate Cartilage. Pediatr Res. 2007;61:267–72. PubMed

Heifetz A, Keenan RW, Elbein AD. Mechanism of action of tunicamycin on the UDP-GlcNAc:dolichyl-phosphate Glc-NAc-1-phosphate transferase. Biochemistry. 1979;18:2186–92. PubMed

Helms JB, Rothman JE. Inhibition by brefeldin a of a Golgi membrane enzyme that catalyses exchange of guanine nucleotide bound to ARF. Nature. 1992;360:352–4. PubMed

Sehgal P, Szalai P, Olesen C, Praetorius HA, Nissen P, Christensen SB, et al. Inhibition of the sarco/endoplasmic reticulum (ER) Ca2+-ATPase by thapsigargin analogs induces cell death via ER Ca2+ depletion and the unfolded protein response. J Biol Chem. 2017;292:19656–73. PubMed PMC

Schröder M, Kaufman RJ. ER stress and the unfolded protein response. Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis. Mutat Res; 2005;569(1–2):29–63 PubMed

Wang L, Perera BGK, Hari SB, Bhhatarai B, Backes BJ, Seeliger MA, et al. Divergent allosteric control of the IRE1α endoribonuclease using kinase inhibitors. Nat Chem Biol. 2012;8:982–9. PubMed PMC

Ghosh R, Wang L, Wang ES, Perera BGK, Igbaria A, Morita S, et al. Allosteric inhibition of the IRE1α RNase preserves cell viability and function during endoplasmic reticulum stress. Cell. 2014;158:534–48. PubMed PMC

Kostas M, Haugsten EM, Zhen Y, Sørensen V, Szybowska P, Fiorito E, et al. Protein Tyrosine Phosphatase Receptor Type G (PTPRG) Controls Fibroblast Growth Factor Receptor (FGFR) 1 Activity and Influences Sensitivity to FGFR Kinase Inhibitors. Mol Cell Proteomics. 2018;17:850–70. PubMed PMC

Marini JC, Forlino A, Bächinger HP, Bishop NJ, Byers PH, De Paepe A, et al. Osteogenesis imperfecta. Nat Rev Dis Primers. 2017;3:17052. 10.1038/nrdp.2017.52. PubMed

Claeys L, Storoni S, Eekhoff M, Elting M, Wisse L, Pals G, et al. Collagen transport and related pathways in Osteogenesis Imperfecta. Hum Genet. 2021;140:1121–41. PubMed PMC

Duran I, Nevarez L, Sarukhanov A, Wu S, Lee K, Krejci P, et al. HSP47 and FKBP65 cooperate in the synthesis of type I procollagen. Hum Mol Genet. 2014;24:1918–28. PubMed PMC

Mirigian LS, Makareeva E, Mertz EL, Omari S, Roberts-Pilgrim AM, Oestreich AK, et al. Osteoblast Malfunction Caused by Cell Stress Response to Procollagen Misfolding in α2(I)-G610C Mouse Model of Osteogenesis Imperfecta. J Bone Miner Res. 2016;31:1608–16. PubMed PMC

Lietman CD, Rajagopal A, Homan EP, Munivez E, Jiang MM, Bertin TK, et al. Connective tissue alterations in Fkbp10-/- mice. Hum Mol Genet. 2014;23:4822–31. PubMed PMC

Lisse TS, Thiele F, Fuchs H, Hans W, Przemeck GKH, Abe K, et al. ER stress-mediated apoptosis in a new mouse model of Osteogenesis imperfecta. PLoS Genet. 2008;4(2):e7. 10.1371/journal.pgen.0040007. PubMed PMC

Duran I, Zieba J, Csukasi F, Martin JH, Wachtell D, Barad M, et al. 4-PBA Treatment Improves Bone Phenotypes in the Aga2 Mouse Model of Osteogenesis Imperfecta. J Bone Miner Res. 2022;37(4):675–86. PubMed PMC

Joeng KS, Schumacher CA, Zylstra-Diegel CR, Long F, Williams BO. Lrp5 and Lrp6 redundantly control skeletal development in the mouse embryo. Dev Biol. 2011;359:222–9. PubMed PMC

Jacob AL, Smith C, Partanen J, Ornitz DM. Fibroblast growth factor receptor 1 signaling in the osteo-chondrogenic cell lineage regulates sequential steps of osteoblast maturation. Dev Biol. 2006;296:315–28. PubMed PMC

Pinson KI, Brennan J, Monkley S, Avery BJ, Skarnes WC. An LDL-receptor-related protein mediates Wnt signalling in mice. Nature. 2000;407:535–8. PubMed

Ishikawa TO, Tamai Y, Zorn AM, Yoshida H, Seldin MF, Nishikawa SI, et al. Mouse Wnt receptor gene Fzd5 is essential for yolk sac and placental angiogenesis. Development. 2001;128:25–33. PubMed

Davidson G, Wu W, Shen J, Bilic J, Fenger U, Stannek P, et al. Casein kinase 1 gamma couples Wnt receptor activation to cytoplasmic signal transduction. Nature. 2005;438:867–72. PubMed

Xu Q, Wang Y, Dabdoub A, Smallwood PM, Williams J, Woods C, et al. Vascular development in the retina and inner ear: Control by Norrin and Frizzled-4, a high-affinity ligand-receptor pair. Cell. 2004;116:883–95. PubMed

Landau DA, Tausch E, Taylor-Weiner AN, Stewart C, Reiter JG, Bahlo J, et al. Mutations driving CLL and their evolution in progression and relapse. Nature. 2015;526:525–30. PubMed PMC

Rosati E, Sabatini R, Rampino G, De Falco F, Di Ianni M, Falzetti F, et al. Novel targets for endoplasmic reticulum stress-induced apoptosis in B-CLL. Blood. 2010;116:2713–23. PubMed

Baskar S, Ka YK, Hofer T, Levy JM, Kennedy MG, Lee E, et al. Unique cell surface expression of receptor tyrosine kinase ROR1 in human B-cell chronic lymphocytic leukemia. Clin Cancer Res. 2008;14:396–404. PubMed

Choi MY, Widhopf GF, Ghia EM, Kidwell RL, Hasan MK, Yu J, et al. Phase I Trial: Cirmtuzumab Inhibits ROR1 Signaling and Stemness Signatures in Patients with Chronic Lymphocytic Leukemia. Cell Stem Cell. 2018;22:951–9. PubMed PMC

Fafilek B, Balek L, Bosakova MK, Varecha M, Nita A, Gregor T, et al. The inositol phosphatase SHIP2 enables sustained ERK activation downstream of FGF receptors by recruiting Src kinases. Sci Signal. 2018;11(548):eaap8608. 10.1126/scisignal.aap8608. PubMed

Gudernova I, Foldynova-Trantirkova S, El Ghannamova B, Fafilek B, Varecha M, Balek L, et al. One reporter for in-cell activity profiling of majority of protein kinase oncogenes. Elife. 2017;6:e21536. 10.7554/eLife.21536. PubMed PMC

Dvorak P, Bednar D, Vanacek P, Balek L, Eiselleova L, Stepankova V, et al. Computer-assisted engineering of hyperstable fibroblast growth factor 2. Biotechnol Bioeng. 2018;115:850–62. PubMed

Harding HP, Novoa I, Zhang Y, Zeng H, Wek R, Schapira M, et al. Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol Cell. 2000;6:1099–108. PubMed

Yoon YM, Lee JH, Yun SP, Han YS, Yun CW, Lee HJ, et al. Tauroursodeoxycholic acid reduces ER stress by regulating of Akt-dependent cellular prion protein. Sci Rep. 2016;6:39838. 10.1038/srep39838. PubMed PMC

Haugsten EM, Sørensen V, Brech A, Olsnes S, Wesche J. Different intracellular trafficking of FGF1 endocytosed by the four homologous FGF receptors. J Cell Sci. 2005;118:3869–81. PubMed

Zakrzewska M, Haugsten EM, Nadratowska-Wesolowska B, Oppelt A, Hausott B, Jin Y, et al. ERK-mediated phosphorylation of fibroblast growth factor receptor 1 on Ser777 inhibits signaling. Sci Signal. 2013;12;6(262):ra11. 10.1126/scisignal.2003087 PubMed

Haugsten EM, Zakrzewska M, Brech A, Pust S, Olsnes S, Sandvig K, et al. Clathrin- and dynamin-independent endocytosis of FGFR3–implications for signalling. PLoS ONE. 2011;6(7):e21708. 10.1371/journal.pone.0021708. PubMed PMC

Annamalai B, Liu X, Gopal U, Isaacs JS. Hsp90 is an essential regulator of EphA2 receptor stability and signaling: Implications for cancer cell migration and metastasis. Mol Cancer Res. 2009;7:1021–32. PubMed PMC

Ou W Bin, Hubert C, Corson JM, Bueno R, Flynn DL, Sugarbaker DJ, et al. Targeted inhibition of multiple receptor tyrosine kinases in mesothelioma. Neoplasia. 2011;13:12–22 PubMed PMC

Laederich MB, Degnin CR, Lunstrum GP, Holden P, Horton WA. Fibroblast Growth Factor Receptor 3 (FGFR3) is a strong Heat shock protein 90 (Hsp90) client: Implications for therapeutic manipulation. J Biol Chem. 2011;286:19597–604. PubMed PMC

Tang CHA, Ranatunga S, Kriss CL, Cubitt CL, Tao J, Pinilla-Ibarz JA, et al. Inhibition of ER stress-associated IRE-1/XBP-1 pathway reduces leukemic cell survival. J Clin Investig. 2014;124:2585–98. PubMed PMC

Matsuo K, Machida H, Frimer M, Marcus JZ, Pejovic T, Roman LD, et al. Prognosis of women with stage I endometrioid endometrial cancer and synchronous stage I endometrioid ovarian cancer. Gynecol Oncol. 2017;147:558–64. PubMed PMC

Lhomond S, Avril T, Dejeans N, Voutetakis K, Doultsinos D, McMahon M, et al. Dual IRE1 RNase functions dictate glioblastoma development. EMBO Mol Med. 2018;15(2): e16731. 10.15252/emmm.202216731. PubMed PMC

Lee AH, Iwakoshi NN, Anderson KC, Glimcher LH. Proteasome inhibitors disrupt the unfolded protein response in myeloma cells. Proc Natl Acad Sci U S A. 2003;100:9946–51. PubMed PMC

Dalton LE, Clarke HJ, Knight J, Lawson MH, Wason J, Lomas DA, et al. The endoplasmic reticulum stress marker CHOP predicts survival in malignant mesothelioma. Br J Cancer. 2013;108:1340–7. PubMed PMC

Chen X, Iliopoulos D, Zhang Q, Tang Q, Greenblatt MB, Hatziapostolou M, et al. XBP1 promotes triple-negative breast cancer by controlling the HIF1α pathway. Nature. 2014;508:103–7. PubMed PMC

Kaucká M, Krejčí P, Plevová K, Pavlová Š, Procházková J, Janovská P, et al. Post-translational modifications regulate signalling by Ror1. Acta Physiol. 2011;203:351–62. PubMed

McKenzie J, Smith C, Karuppaiah K, Langberg J, Silva MJ, Ornitz DM. Osteocyte Death and Bone Overgrowth in Mice Lacking Fibroblast Growth Factor Receptors 1 and 2 in Mature Osteoblasts and Osteocytes. J Bone Miner Res. 2019;34:1660–75. PubMed PMC

Roschger P, Fratzl-Zelman N, Misof BM, Glorieux FH, Klaushofer K, Rauch F. Evidence that abnormal high bone mineralization in growing children with osteogenesis imperfecta is not associated with specific collagen mutations. Calcif Tissue Int. 2008;82:263–70. PubMed

Forlino A, Marini JC. Osteogenesis imperfecta. Lancet. 2016;387(10028):1657–71. PubMed PMC

Houschyar KS, Tapking C, Borrelli MR, Popp D, Duscher D, Maan ZN, et al. Wnt Pathway in Bone Repair and Regeneration - What Do We Know So Far. Front Cell Dev Biol. 2019;6:170. 10.3389/fcell.2018.00170. PubMed PMC

Laine CM, Joeng KS, Campeau PM, Kiviranta R, Tarkkonen K, Grover M, et al. WNT1 mutations in early-onset osteoporosis and osteogenesis imperfecta. N Engl J Med. 2013;368:1809–16. PubMed PMC

Gong Y, Slee RB, Fukai N, Rawadi G, Roman-Roman S, Reginato AM, et al. LDL receptor-related protein 5 (LRP5) affects bone accrual and eye development. Cell. 2001;107:513–23. PubMed

Kalajzic I, Terzic J, Rumboldt Z, Mack K, Naprta A, Ledgard F, et al. Osteoblastic response to the defective matrix in the osteogenesis imperfecta murine (oim) mouse. Endocrinology. 2002;143:1594–601. PubMed

Gioia R, Panaroni C, Besio R, Palladini G, Merlini G, Giansanti V, et al. Impaired osteoblastogenesis in a murine model of dominant osteogenesis imperfecta: a new target for osteogenesis imperfecta pharmacological therapy. Stem Cells. 2012;30:1465–76. PubMed PMC

Lombardi G, Di Somma C, Rubino M, Faggiano A, Vuolo L, Guerra E, et al. The roles of parathyroid hormone in bone remodeling: prospects for novel therapeutics. J Endocrinol Invest. 2011;34:18–22. PubMed

Kimura T, Bosakova M, Nonaka Y, Hruba E, Yasuda K, Futakawa S, et al. An RNA aptamer restores defective bone growth in FGFR3-related skeletal dysplasia in mice. Sci Transl Med. 2021;13(592):eaba4226. 10.1126/scitranslmed.aba4226. PubMed

Mossine VV, Waters JK, Hannink M, Mawhinney TP. PiggyBac transposon plus insulators overcome epigenetic silencing to provide for stable signaling pathway reporter cell lines. PLoS ONE. 2013;8(12):e85494. 10.1371/journal.pone.0085494. PubMed PMC

Fafilek B, Hampl M, Ricankova N, Vesela I, Balek L, Kunova Bosakova M, et al. Statins do not inhibit the FGFR signaling in chondrocytes. Osteoarthritis Cartilage. 2017;25:1522–30. PubMed