Hyper-IL-6
{{Short description|Designer cytokine}}
{{COI|date=June 2020}}
{{Orphan|date=June 2020}}
Hyper-IL-6 is a designer cytokine, which was generated by the German biochemist Stefan Rose-John.{{cite journal |last1=Fischer |first1=Martina |last2=Goldschmitt |first2=Jutta |last3=Peschel |first3=Christian |last4=Brakenhoff |first4=Just P. G. |last5=Kallen |first5=Karl-Josef |last6=Wollmer |first6=Axel |last7=Grötzinger |first7=Joachim |last8=Rose-John |first8=Stefan |title=A bioactive designer cytokine for human hematopoietic progenitor cell expansion |journal=Nature Biotechnology |date=February 1997 |volume=15 |issue=2 |pages=142–145 |doi=10.1038/nbt0297-142 |pmid=9035138 |s2cid=22721071 }} Hyper-IL-6 is a fusion protein of the four-helical cytokine Interleukin-6 and the soluble Interleukin-6 receptor which are covalently linked by a flexible peptide linker. Interleukin-6 on target cells binds to a membrane bound Interleukin-6 receptor.{{cite journal |last1=Jones |first1=Simon A. |last2=Jenkins |first2=Brendan J. |title=Recent insights into targeting the IL-6 cytokine family in inflammatory diseases and cancer |journal=Nature Reviews Immunology |date=25 September 2018 |volume=18 |issue=12 |pages=773–789 |doi=10.1038/s41577-018-0066-7 |pmid=30254251 |s2cid=52823385 |url=http://orca.cf.ac.uk/116367/1/NRI-2018-accepted%20version.pdf }} The complex of Interleukin-6 and the Interleukin-6 receptor associate with a second receptor protein called gp130, which dimerises and initiates intracellular signal transduction.{{cite journal |last1=Schaper |first1=Fred |last2=Rose-John |first2=Stefan |title=Interleukin-6: Biology, signaling and strategies of blockade |journal=Cytokine & Growth Factor Reviews |date=October 2015 |volume=26 |issue=5 |pages=475–487 |doi=10.1016/j.cytogfr.2015.07.004 |pmid=26189695 }} Gp130 is expressed on all cells of the human body whereas the Interleukin-6 receptor is only found on few cells such as hepatocytes and some leukocytes.{{cite journal |last1=Taga |first1=Tetsuya |last2=Kishimoto |first2=Tadamitsu |title=Gp130 and the interleukin-6 Family of Cytokines |journal=Annual Review of Immunology |date=April 1997 |volume=15 |issue=1 |pages=797–819 |doi=10.1146/annurev.immunol.15.1.797 |pmid=9143707 }} Neither Interleukin-6 nor the Interleukin-6 receptor have a measurable affinity for gp130.{{cite journal |last1=Rose-John |first1=Stefan |title=IL-6 Trans-Signaling via the Soluble IL-6 Receptor: Importance for the Pro-Inflammatory Activities of IL-6 |journal=International Journal of Biological Sciences |date=2012 |volume=8 |issue=9 |pages=1237–1247 |doi=10.7150/ijbs.4989 |pmid=23136552 |pmc=3491447 }} Therefore, cells, which only express gp130 but no Interleukin-6 receptor are not responsive to Interleukin-6. It was found, however, that the membrane-bound Interleukin-6 receptor can be cleaved from the cell membrane generating a soluble Interleukin-6 receptor.{{cite journal |last1=Mülberg |first1=Jürgen |last2=Schooltink |first2=Heidi |last3=Stoyan |first3=Tanja |last4=Günther |first4=Monika |last5=Graeve |first5=Lutz |last6=Buse |first6=Gerhard |last7=Mackiewicz |first7=Andrzej |last8=Heinrich |first8=Peter C. |last9=Rose-John |first9=Stefan |title=The soluble interleukin-6 receptor is generated by shedding |journal=European Journal of Immunology |date=February 1993 |volume=23 |issue=2 |pages=473–480 |doi=10.1002/eji.1830230226 |pmid=8436181 |s2cid=22834660 }} The soluble Interleukin-6 receptor can bind the ligand Interleukin-6 with similar affinity as the membrane-bound Interleukin-6 receptor and the complex of Interleukin-6 and the soluble Interleukin-6 receptor can bind to gp130 on cells, which only express gp130 but no Interleukin-6 receptor.{{cite journal |last1=Mackiewicz |first1=A. |last2=Schooltink |first2=H. |last3=Heinrich |first3=P. C. |last4=Rose-John |first4=S. |title=Complex of soluble human IL-6-receptor/IL-6 up-regulates expression of acute-phase proteins. |journal=The Journal of Immunology |date=15 September 1992 |volume=149 |issue=6 |pages=2021–2027 |doi=10.4049/jimmunol.149.6.2021 |pmid=1381393 |url=http://www.jimmunol.org/cgi/pmidlookup?view=long&pmid=1381393 }} The mode of signaling via the soluble Interleukin-6 receptor has been named Interleukin-6 trans-signaling whereas Interleukin-6 signaling via the membrane-bound Interleukin-6 receptor is referred to as Interleukin-6 classic signaling.{{cite journal |last1=Rose-John |first1=S |last2=Heinrich |first2=P C |title=Soluble receptors for cytokines and growth factors: generation and biological function |journal=Biochemical Journal |date=1 June 1994 |volume=300 |issue=2 |pages=281–290 |doi=10.1042/bj3000281 |pmid=8002928 |pmc=1138158 }} Therefore, the generation of the soluble Interleukin-6 receptor enables cells to respond to Interleukin-6, which in the absence of soluble Interleukin-6 receptor would be completely unresponsive to the cytokine.
Molecular construction of Hyper-IL-6
In order to generate a molecular tool to discriminate between Interleukin-6 classic signaling and Interleukin-6 trans-signaling, a cDNA coding for human Interleukin-6 and a cDNA coding for the human soluble Interleukin-6 receptor were connected by a cDNA coding for a 13 amino acids long linker, which was long enough to bridge the 40 Å distance between the COOH terminus of the soluble Interleukin-6 receptor and the NH2 terminus of human Interleukin-6.{{cite journal |last1=Grotzinger |first1=Joachim |last2=Kurapkat |first2=Günther |last3=Wollmer |first3=Axel |last4=Kalai |first4=Michael |last5=Rose-John |first5=Stefan |title=The family of the IL-6-Type cytokines: Specificity and promiscuity of the receptor complexes |journal=Proteins: Structure, Function, and Genetics |date=January 1997 |volume=27 |issue=1 |pages=96–109 |doi=10.1002/(SICI)1097-0134(199701)27:1<96::AID-PROT10>3.0.CO;2-D |pmid=9037715 |s2cid=35512559 }} The generated cDNA was expressed in yeast cells and in mammalian cells and it was shown that.{{cite journal |last1=Peters |first1=Malte |last2=Müller |first2=Albrecht M. |last3=Rose-John |first3=Stefan |title=Interleukin-6 and Soluble Interleukin-6 Receptor: Direct Stimulation of gp130 and Hematopoiesis |journal=Blood |date=15 November 1998 |volume=92 |issue=10 |pages=3495–3504 |doi=10.1182/blood.V92.10.3495 |pmid=9808540 }}
Use of Hyper-IL-6 to analyse IL-6 signaling
Hyper-IL-6 has been used to test which cells depend on Interleukin-6 trans-signaling in their response to the cytokine Interleukin-6. To this end, cells were treated with Interleukin-6 and alternatively with Hyper-IL-6. Cells, which respond to Interleukin-6 alone do express an Interleukin-6 receptor whereas cells, which only respond to Hyper-IL-6 but not to Interleukin-6 alone depend in their response to the cytokine on Interleukin-6 trans-signaling.{{cite journal |last1=Wolf |first1=Janina |last2=Rose-John |first2=Stefan |last3=Garbers |first3=Christoph |title=Interleukin-6 and its receptors: A highly regulated and dynamic system |journal=Cytokine |date=November 2014 |volume=70 |issue=1 |pages=11–20 |doi=10.1016/j.cyto.2014.05.024 |pmid=24986424 }} It turned out that hematopoietic stem cells,{{cite journal |last1=Audet |first1=Julie |last2=Miller |first2=Cindy L. |last3=Rose-John |first3=Stefan |last4=Piret |first4=James M. |last5=Eaves |first5=Connie J. |title=Distinct role of gp130 activation in promoting self-renewal divisions by mitogenically stimulated murine hematopoietic stem cells |journal=Proceedings of the National Academy of Sciences of the United States of America |date=13 February 2001 |volume=98 |issue=4 |pages=1757–1762 |doi=10.1073/pnas.98.4.1757 |pmid=11172024 |pmc=29330 |jstor=3054949 |bibcode=2001PNAS...98.1757A |doi-access=free }} neural cells,{{cite journal |last1=März |first1=Pia |last2=Cheng |first2=Jr-Gang |last3=Gadient |first3=Reto A. |last4=Patterson |first4=Paul H. |last5=Stoyan |first5=Tanja |last6=Otten |first6=Uwe |last7=Rose-John |first7=Stefan |title=Sympathetic neurons can produce and respond to interleukin 6 |journal=Proceedings of the National Academy of Sciences of the United States of America |date=17 March 1998 |volume=95 |issue=6 |pages=3251–3256 |doi=10.1073/pnas.95.6.3251 |pmid=9501249 |pmc=19728 |jstor=44831 |bibcode=1998PNAS...95.3251M |doi-access=free }} smooth muscle cells{{cite journal |last1=Klouche |first1=Mariam |last2=Bhakdi |first2=Sucharit |last3=Hemmes |first3=Monika |last4=Rose-John |first4=Stefan |title=Novel Path to Activation of Vascular Smooth Muscle Cells: Up-Regulation of gp130 Creates an Autocrine Activation Loop by IL-6 and Its Soluble Receptor |journal=The Journal of Immunology |date=15 October 1999 |volume=163 |issue=8 |pages=4583–4589 |doi=10.4049/jimmunol.163.8.4583 |pmid=10510402 |url=http://www.jimmunol.org/cgi/pmidlookup?view=long&pmid=10510402 }} and endothelial cells{{cite journal |last1=Romano |first1=Maria |last2=Sironi |first2=Marina |last3=Toniatti |first3=Carlo |last4=Polentarutti |first4=Nadia |last5=Fruscella |first5=Paolo |last6=Ghezzi |first6=Pietro |last7=Faggioni |first7=Raffaella |last8=Luini |first8=Walter |last9=van Hinsbergh |first9=Victor |last10=Sozzani |first10=Silvano |last11=Bussolino |first11=Federico |last12=Poli |first12=Valeria |last13=Ciliberto |first13=Gennaro |last14=Mantovani |first14=Alberto |title=Role of IL-6 and Its Soluble Receptor in Induction of Chemokines and Leukocyte Recruitment |journal=Immunity |date=March 1997 |volume=6 |issue=3 |pages=315–325 |doi=10.1016/s1074-7613(00)80334-9 |pmid=9075932 |doi-access=free }} are typical target cells of Interleukin-6 trans-signaling.
The concept of Interleukin-6 trans-signaling
The Hyper-IL-6 protein has also been used to explore the physiologic role of Interleukin-6 trans-signaling in vivo. It turned out that this signaling mode was involved in many types of inflammation{{cite journal |last1=Calabrese |first1=Leonard H. |last2=Rose-John |first2=Stefan |title=IL-6 biology: implications for clinical targeting in rheumatic disease |journal=Nature Reviews Rheumatology |date=19 August 2014 |volume=10 |issue=12 |pages=720–727 |doi=10.1038/nrrheum.2014.127 |pmid=25136784 |s2cid=31635688 |doi-access=free }} and cancer.{{cite journal |last1=Schmidt |first1=Stefanie |last2=Schumacher |first2=Neele |last3=Schwarz |first3=Jeanette |last4=Tangermann |first4=Simone |last5=Kenner |first5=Lukas |last6=Schlederer |first6=Michaela |last7=Sibilia |first7=Maria |last8=Linder |first8=Markus |last9=Altendorf-Hofmann |first9=Annelore |last10=Knösel |first10=Thomas |last11=Gruber |first11=Elisabeth S. |last12=Oberhuber |first12=Georg |last13=Bolik |first13=Julia |last14=Rehman |first14=Ateequr |last15=Sinha |first15=Anupam |last16=Lokau |first16=Juliane |last17=Arnold |first17=Philipp |last18=Cabron |first18=Anne-Sophie |last19=Zunke |first19=Friederike |last20=Becker-Pauly |first20=Christoph |last21=Preaudet |first21=Adele |last22=Nguyen |first22=Paul |last23=Huynh |first23=Jennifer |last24=Afshar-Sterle |first24=Shoukat |last25=Chand |first25=Ashwini L. |last26=Westermann |first26=Jürgen |last27=Dempsey |first27=Peter J. |last28=Garbers |first28=Christoph |last29=Schmidt-Arras |first29=Dirk |last30=Rosenstiel |first30=Philip |last31=Putoczki |first31=Tracy |last32=Ernst |first32=Matthias |last33=Rose-John |first33=Stefan |title=ADAM17 is required for EGF-R–induced intestinal tumors via IL-6 trans-signaling |journal=Journal of Experimental Medicine |date=2 April 2018 |volume=215 |issue=4 |pages=1205–1225 |doi=10.1084/jem.20171696 |pmid=29472497 |pmc=5881468 }}
Hyper-IL-6 has helped to establish the concept of Interleukin-6 trans-signaling.{{cite journal |last1=Rose-John |first1=Stefan |last2=Winthrop |first2=Kevin |last3=Calabrese |first3=Leonard |title=The role of IL-6 in host defence against infections: immunobiology and clinical implications |journal=Nature Reviews Rheumatology |date=15 June 2017 |volume=13 |issue=7 |pages=399–409 |doi=10.1038/nrrheum.2017.83 |pmid=28615731 |s2cid=205519501 }} Interleukin-6 trans-signaling mediates the pro-inflammatory activities of Interleukin-6 whereas Interleukin-6 classic signaling governs the protective and regenerative Interleukin-6 activities.{{cite journal |last1=Garbers |first1=Christoph |last2=Heink |first2=Sylvia |last3=Korn |first3=Thomas |last4=Rose-John |first4=Stefan |title=Interleukin-6: designing specific therapeutics for a complex cytokine |journal=Nature Reviews Drug Discovery |date=4 May 2018 |volume=17 |issue=6 |pages=395–412 |doi=10.1038/nrd.2018.45 |pmid=29725131 |s2cid=19238101 }} Recently, in breast cancer patients, it was shown with the help of Hyper-IL-6 that IL-6 trans-signaling via phosphoinositid-3-kinase signaling activates disseminated cancer cells long before metastases are formed.Werner-Klein M, Grujovic A, Irlbeck C, Obradović M, Hoffmann M, Koerkel-Qu H, Lu X, Treitschke S, Köstler C, Botteron C, Weidele K, Werno C, Polzer B, Kirsch S, Gužvić M, Warfsmann J, Honarnejad K, Czyz Z, Feliciello G, Blochberger I, Grunewald S, Schneider E, Haunschild G, Patwary N, Guetter S, Huber S, Rack B, Harbeck N, Buchholz S, Rümmele P, Heine N, Rose-John S, Klein CA (2020) Interleukin-6 trans-signaling is a candidate mechanism to drive progression of human DCCs during clinical latency. Nat Commun 11(1):4977 In addition, it was demonstrated in mice that Hyper-IL-6 transneuronal delivery enabled functional recovery after severe spinal cord injury.Leibinger M, Zeitler C, Gobrecht P, Anastasia Andreadaki A, Gisselmann G, Fischer D (2021) Transneuronal delivery of hyper-interleukin-6 enables functional recovery after severe spinal cord injury in mice. Nat Commun 12(1):391