US010106603B2
`
`( 12 ) United States Patent
`Cook et al .
`
`( 10 ) Patent No . : US 10 , 106 , 603 B2
`( 45 ) Date of Patent :
`Oct . 23 , 2018
`
`( 54 ) TREATMENT OF FIBROSIS
`( 71 ) Applicants : Singapore Health Services PTE LTD . ,
`Singapore ( SG ) ; National University of
`Singapore , Singapore ( SG )
`( 72 ) Inventors : Stuart Alexander Cook , Singapore
`( SG ) ; Sebastian Schaefer , Singapore
`( SG )
`( 73 ) Assignees : Singapore Health Services PTE LTD ,
`Singapore ( SG ) ; National University of
`Singapore , Singapore ( SG )
`Subject to any disclaimer , the term of this
`patent is extended or adjusted under 35
`U . S . C . 154 ( b ) by 0 days .
`( 21 ) Appl . No . : 15 / 988 , 463
`May 24 , 2018
`( 22 )
`Filed :
`Prior Publication Data
`( 65 )
`US 2018 / 0265579 A1 Sep . 20 , 2018
`
`( * ) Notice :
`
`Related U . S . Application Data
`( 62 ) Division of application No . 15 / 381 , 622 , filed on Dec .
`16 , 2016 , now Pat . No . 10 , 035 , 852 .
`Foreign Application Priority Data
`( 30 )
`Dec . 16 , 2015
`( GB ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1522186 . 4
`( 51 ) Int . Ci .
`A61K 39 / 395
`COOK 16 / 24
`A61K 39 / 00
`( 52 ) U . S . CI .
`CPC . . . . . . CO7K 16 / 244 ( 2013 . 01 ) ; A61K 2039 / 505
`( 2013 . 01 ) ; A61K 2039 / 54 ( 2013 . 01 ) ; COOK
`2317 / 20 ( 2013 . 01 ) ; CO7K 2317 / 76 ( 2013 . 01 )
`( 58 ) Field of Classification Search
`. . . . A61K 2039 / 505
`CPC . . . . . . . . . . .
`See application file for complete search history .
`
`( 2006 . 01 )
`( 2006 . 01 )
`( 2006 . 01 )
`
`( 56 )
`
`References Cited
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`KR
`wo
`Wo
`WO
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`WO
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`OTHER PUBLICATIONS
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`( Continued )
`
`Primary Examiner — Prema M Mertz
`( 74 ) Attorney , Agent , or Firm — Wolf , Greenfield &
`Sacks , P . C .
`
`ABSTRACT
`( 57 )
`Aspects of the disclosure relate to the treatment , prevention
`or alleviation of conditions such as fibrosis in a subject . In
`some embodiments , the treatment , prevention or alleviation
`of fibrosis in a subject through the administration of an agent
`capable of inhibiting the action of Interleukin 11 ( IL - 11 ) is
`disclosed .
`
`10 Claims , 66 Drawing Sheets
`Specification includes a Sequence Listing .
`
`Lassen - Exhibit 1001, p. 1
`
`

`

`US 10 , 106 , 603 B2
`Page 2
`
`( 56 )
`
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`OTHER PUBLICATIONS
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`lung fibrosis and epithelial damage in a humanized SCID idiopathic
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`during gastrointestinal tumorigenesis and can be targeted therapeu
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`ccr . 2013 . 06 . 017 .
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`M111 . 349167 .
`
`Lassen - Exhibit 1001, p. 2
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 1 of 66
`
`US 10 , 106 , 603 B2
`
`* * *
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`
`L
`
`Figure 1A
`
`Figure 1C
`
`Lassen - Exhibit 1001, p. 3
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`Sheet 2 of 66
`
`US 10 , 106 , 603 B2
`
`Figure 2B
`
`W
`
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`
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`
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`
`Figure 20
`
`Lassen - Exhibit 1001, p. 4
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 2
`
`Oct . 23 , 2018
`
`Sheet 3 of 66
`
`US 10 , 106 , 603 B2
`
`
`
`PCS BOLA
`
`* * * * * *
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`Figure 3B
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`88
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`+ 1 +
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`Lassen - Exhibit 1001, p. 5
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 4 of 66
`
`US 10 , 106 , 603 B2
`
`Figure 30
`
`Figure 3D
`
`Lassen - Exhibit 1001, p. 6
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 5 of 66
`
`US 10 , 106 , 603 B2
`
`. 4 . 4 . 4 . 5 . 6 . 6 . 0 . 1
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`
`TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
`
`Lassen - Exhibit 1001, p. 7
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 6 of 66
`
`US 10 , 106 , 603 B2
`
`?t ?xn811
`
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`
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`
`Lassen - Exhibit 1001, p. 8
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 7 of 66
`
`US 10 , 106 , 603 B2
`
`Figure 5B
`
`Figure 5D
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`Lassen - Exhibit 1001, p. 9
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 8 of 66
`
`US 10 , 106 , 603 B2
`
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`
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`Lassen - Exhibit 1001, p. 10
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 9 of 66
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`US 10 , 106 , 603 B2
`
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`Lassen - Exhibit 1001, p. 11
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 10 of 66
`
`US 10 , 106 , 603 B2
`
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`Lassen - Exhibit 1001, p. 12
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`Sheet 11 of 66
`
`US 10 , 106 , 603 B2
`
`Figure 8B
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`Lassen - Exhibit 1001, p. 13
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 12 of 66
`
`US 10 , 106 , 603 B2
`
`Kidney
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`
`Lassen - Exhibit 1001, p. 14
`
`

`

`U . S . Patent
`
`
`
`Tissue Injury
`
`21
`
`2838
`
`Opholog1010101010101010100101011000000101010
`
`Oct . 23 , 2018
`
`Sheet 13 of 66
`
`US 10 , 106 , 603 B2
`
`* * *
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`Lassen - Exhibit 1001, p. 15
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 14 of 66
`
`US 10 , 106 , 603 B2
`
`
`
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`
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`Lassen - Exhibit 1001, p. 16
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 15 of 66
`
`US 10 , 106 , 603 B2
`
`mmmm .
`
`ACTGCCGCGGCCCTGCTGCICAGGGCACATGCCTCCCCTCCCCAGGCCGCGCCCCAGCIGACCCICGGGG
`CICCCCCGGCAGCGGACAGGGAAGGGTTAAAGGCCCCCGGCTCCCTGCCCCCTGCCCIGGGGAACCCCIG
`GCCCTGTGGGGACATGAACTGTGTTTGCCGCCTGGTCCTGGTCGTGCIGAGCCTGTGGCCAGATACAGCT
`GTCGCCCCTGGGCCACCACCTGGCCCCCCTCGAGTITCOCCAGACCCICGGGCCGAGCIGGACAGCACCG
`TGCICCTGACCCGCICICICCIGGCGGACACGCGGCAGCTGGCTGCACAGCIGAGGGACAAATTCCCAGO
`TGACGGGGACCACAACCTGGATTCCCTGCCCACCCTGGCCATGAGTGCGGGGGCACTGGGAGCTCTACAG
`CICCCAGGTGIGCTGACAAGGCTGCGAGCGGACCTACTGTCCTACCTGCGGCACGTGCAGTGGCTGCGCC
`GGGCAGGTGGCTCTTCCCTGAAGACCCTGGAGCCCGAGCTGGGCACCCTGCAGGCCCGACTGGACCGGCT
`GCTGCGCCGGCTGCAGCICCIGATGTOCCGCCTGGCCCTGCCCCAGCCACCCCCGGACCCGCCGGCGCCO
`CCGCTGGCGCCCCCCTCCTCAGCCIGGGGGGGCATCAGGGCCGCCCACGCCATCCTGGGGGGGCTGCACC
`TGACACTIGACIGGGCCGTGAGGGGACTGCTGCTGCTGAAGACTCGGCIGTGACCCGGGGCCCAAAGCCA
`CCACCGTBOLO AANGOUAGA OODATTTATTTATTTATTICAGTACTGGGGGCGAAACAGCCAGGIGA
`TCCCCCCGCCATTATCTCCCCCTAGTTAGAGACAGTCCTTCCGTGAGGCCTGGGGGGCATCTGTGCCTTA
`TITATACTTATITATTICAGGAGCAGGGGTGGGAGGCAGGTGGACTCCTGGGTCCCCGAGGAGGAGGGGA
`CIGGGGTCCCGGATTCTTGGGICTCCAAGAAGTCTGTCCACAGACTTCTGCCCIGGCICTTCCCCAICTA
`GECSKEGGUAEGLAGAHAR TATTTATTTAAGCAATTACTTTTCATGTTGGGGTGGGGACGGAGGGGAA
`AGGGAAGCCTGGGTTITIGTACAAAAATGTGAGAAACCITIGTGAGACAGAGAACAGGGAATTAAATGIG
`TCATACATATCCACTTGAGGGCGATTTGTCTGAGAGCTGGGGCIGGATGCTTGGGTAACTGGGGCAGGGO
`AGGTGGAGGGGAGACCTCCATICAGGTGGAGGICCCGAGIGGGCGGGGCAGCGACTGGGAGATGGGICGG
`ICACCCAGACAGCICTGIGGAGGCAGGGTCTGAGCCTIGCCTGGGGCCCCGCACIGCATAGGGCCTTTTG
`GAGGCAATCTGA GGTCAC
`GUICIGIT G
`TY 11 AY !
`CAACCTCCACCICCCGGGTTCAAGCAATTCTCCTGCCTCAGCCPCCCGATTAGCTGGGATCACAGGIGTG
`CACCACCATGCCCAGCTAATTATTTATTTCTTTTGTATTTTTAGTAGAGACAGGGTTTCACCATGTTGGC
`CAGGCTGGITTCGAACICCIGACCTCAGGTGATCCTCCTGCCICGGCCTCCCAAAGTGCIGGGATTACAG
`GTGTGAGCCACCACACCTGACCCATAGGTCTTCAATAAATATTTAATGGAAGGTTCCACAAGTCACCCTG
`TGATCAACAGTACCCGTATGGGACAÄAGCTGCAAGGICAAGATOG MANGOCHRXACCATAG
`CAAACIGGAAACAATCTAGATATCCAACAGTGAGGGTTAAGCAACATGGTGCATCTGTGGATAGAACGCC
`ACCCAGCCGCCCGGAGCAGGGACIGTCATICAGGGAGGOTAAGGAGAGAGGCTTGCTTGGGATATAGAAA
`GATATCCTGACATTGGCCAGGCATGGTGGCTCACGCCTGTAATCCTGGCACTTTGGGAGGACGAAGCGAG
`TGGATCACTGAAGTCCAAGAGITCGAGACCGGCCIGCGAGACAIGGCAAAACCCTGICICAAAAAAGAAA
`GAATGATGTCCTGACATGAAACAGCAGGCTACAAAACCACTGCATCCTGTGATCCCAATTTTGTGTTTTT
`CTTTCTATATATGGATTAAAACAAAAATCCTAAAGGGAAATACGCCAAAATGTTGACAATGACIGICICO
`AGGTCAAAGGAGAGAGGTGGGATTGTGGGTGACTTTTAATGTGTATGATTGTCTGTATTTTACAGAATIT
`CTGCCAIGACIGTGTATTTIGCATGACACATTTTAAAAATAATAAACACTATTITTAGAATAACAGAAAA
`( SEO ID NO : 11
`CCTTCCAAAGCCAGATCTT ( SEQ ID NO : 21
`GCCTGGGCAGGAACATATA ( SED ID NO : 3 )
`CCTGGGCAGGAACATATAT ( SEQ ID NO : 4 ]
`GETTCATTATGGCTGTGTT [ SEO ID NO : 51
`
`e 11
`
`Lassen - Exhibit 1001, p. 17
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 16 of 66
`
`US 10 , 106 , 603 B2
`
`* *
`
`* *
`
`IKKIL
`
`UVYVINVIVILLILLLLLL
`
`horrory vybrydory vybrydororoorderprooroordoor
`
`order to order torr
`
`GCIGPAGCTGGTGAGAGGAAGTCCIAGAGGCTATGGACACICTGCIGCIGGGAICACCGAGATGAGCAGO
`AGCIGCICAGGGCIGAGCAGGGTCCTGGTGGCCGIGGOTACAGCCCIGGTGTCTGCCTCCICCCCCIGCC
`CCCAGGCCTGGGGOCCCCCAGGGGTCCAGTATGGGCAGCCAGGGAGGTCCGTGAAGCTGTGTTGTCCTGG
`AGTGACTCCCGGGGACCCAGTGTCCTGGTTICCGGATGGGGAGCCAAAGCTGCTCCAGGGACCIGACICT
`GGGCIAGGGCAIGAACTGGICCIGGCCCAGGCAGACAGCACTGATGAGGGCACCTACATCIGCCAGACCC
`TGGATGGTGCACTTGGGGGCACAGTGACCCTGCAGCIGGGCTACCCICCACCCCGCCCIGTIGTCTCCTG
`CCAAGCAGCCGACTATGAGAACTTCTCTTGCACTTGGAGTCCCAGCCAGATCAGCGGTTTACCCACCCGC
`TACCICACCTCCTACAGGAAGAAGACAGTCCTAGGAGCIGATAGCCAGAGGAGGAGTCCATCCACAGGGO
`CCIGGCCATGCCCACAGGATCCCCTAGGGGCTGCCCGCIGTGTTGICCACGGGGCTGAGITCIGGAGCCA
`GTACCGGATPAAIGTGACTGAGGIGAACCCACTGGGIGCCAGCACACGCCIGCTGGATGIGAGCTTGCAG
`AGCATCTTGCGCCCTGACCCACCCCAGGGCCTGCGGGPAGAGTCAGTACCAGGTTACCCCCGACGCCTGC
`GAGCCAGCTGGACATACCCTGCCTCCTGGCCGTGCCAGCOCCACITCCTGCTCAAGTTCCSITTGCAGTA
`CCGTCCGGCGCAGCATOCAGCCTGGTCCACGGTGGAGCCAGCIGGACTGGAGGAGGTGATCACAGATGCI
`GIGGCIGGGCIGCCCCATGCTGIACGAGICAGTGCCCGGGACITICTAGAIGCIGGCACCIGGAGCACCI
`GGAGCCCGGAGGCCTGGGGAACTCCGAGCACTGE
`ACCAGCATGGGGCCAGCT
`CAGGIGG
`ACACACGCACCCAGAGGIG
`CCTV
`! " VIVIA ILIOHUDHULI
`TCCTGGGACTGGIGGCTGGGGCCCTGGCACIGGGGCTCTGGCTGAGGCIGAGACGGGGIGGGAAGGATGG
`ATCCCCAAAGCCTGGGTTCTTGGCCTCAGTGATTCCAGTGGACAGGCGTCCAGGAGCTCCAAACCTGTAG
`AGGACCCAGGAGGGCTTCGGCAGATTCCACCIATAATTCTGTCTTGCTGGIGTGGATAGAAACCAGO
`A
`TGGITGGATCICAGCOGGAAGIICIGTITGGAGCCCATTICIGIGAGACCCIGTA
`TTTCAAATTIGCAGCTGAAAGGIGCITGIACCICIGATITCACCCCAGAGTTGGAGTICIAS
`STGTGTACATCTGTGTCCATGTGTGACCATGTGTCTGTGAAGGCCAGGGAACATGTATTCCT
`C
`TTCCCTIGCAGGGTIG
`CPGO ATGCATGTATGTAGGTG
`TiTAO
`GAGIGTG
`GTC
`1
`( SEQ ID NO : 6 )
`IGCAGGIGTGAATAAA
`
`y
`
`yyyyyyyyyyyyyyyyyyyhhhhhhhhhhhhhhhhhhhh ththith
`
`Triin
`
`TITUTO
`
`?????
`
`h
`
`S
`
`GGACCATACCAAAGGAGAT ( SEO ID NO : 7 )
`GCGTCTTTGGGAATCCTTT [ SEQ ID NO : 8 )
`GCAGGACAGTAGATCCCT
`( SEO ID NO : 91
`
`GCTCAAGGAACGTGTGTAA ( SEQ ID NO : 101
`
`Figure 12
`
`Lassen - Exhibit 1001, p. 18
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 17 of 66
`
`US 10 , 106 , 603 B2
`
`SEQ ID NO :
`
`12
`
`
`
`siRNA sequence ( 593 )
`
`siRNA 1 AAGAUCUGGCUUUGGAAGGATIT
`CCUUCCAAAGCCAGAUCUUSTAT
`
`GCCUGGGCAGGAACAUAUALTAT
`SiRNA 2 UAUAUGUUCCUGCCCAGGCDTIT
`
`CCUGGGCAGGAACAUAUAUITAT
`
`AUAUAUGUUCCUGCCCAGGSTAT
`
`SiRNA 3
`
`ACCESSION
`
`NM 000641 . 3
`
`NM 0006413
`
`NM 000641 . 3
`
`IL - 11
`
`GGUUCAUUAUGGCUGUGUUSTAT
`SiRNA 4 AACACAGCCAUAAUGAACCITAT
`
`NM 0006413
`
`Figure 13
`
`Lassen - Exhibit 1001, p. 19
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 18 of 66
`
`US 10 , 106 , 603 B2
`
`SEQ I NO :
`
`15
`
`16
`
`
`
`SiRNA sCuence ( 33 )
`
`GGACCAUACCAAAGGAGAULTIT
`SiRNAS AUCUCCUUUGGUAUGGUCCITAT
`
`GCGUCUUUGGGAAUCCUUUSTOT
`
`AAAGGAUUCCCAAAGACGCDTIT
`
`SiRNA 6
`
`GCAGGACAGUAGAUCCCUANTAT
`
`UAGGGAUCUACUGUCCUCCITAT
`
`siRNA 7
`
`GCUCAAGGAACGUGUGUAASTAT
`
`siRNAS UUACACACGUUCCUUGAGCITAT
`
`Figure 14
`
`032324 . 1
`
`U32324 . 1
`
`U32324 . 1
`
`U32324 . 1
`
`IL - IR
`
`IL - IIR
`
`IL - UIR
`
`IL - IR
`
`Lassen - Exhibit 1001, p. 20
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 19 of 66
`
`US 10 , 106 , 603 B2
`
`????? Figure 15
`
`Lassen - Exhibit 1001, p. 21
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 20 of 66
`
`US 10 , 106 , 603 B2
`
`200
`
`160
`Reads ( M )
`
`Total Reads
`mapped
`unique
`
`.
`
`.
`
`cocoon
`
`Wooooooo
`wo
`
`00000000000000
`accoccoccocco
`Woooooooooooo
`Www
`
`Figure 16
`
`PECAM1 - CD31
`
`. . . .
`
`. .
`
`. . . . . . .
`
`. . . . . . . . . .
`
`. . .
`
`. .
`
`. .
`
`. .
`
`. .
`
`. .
`
`. . . .
`
`.
`
`FPKM
`
`100004
`
`Figure 17A
`
`Lassen - Exhibit 1001, p. 22
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`US 10 , 106 , 603 B2
`
`Sheet 21 of 6
`
`Sheet 21 of 66
`
`MYH6
`
`20
`
`WWWWWWWWWWWWWWWWWWWWWW
`
`Atrium
`
`1500m
`
`Figure 17B
`
`TNNT2
`
`I
`
`1994
`
`Figure 170
`
`Lassen - Exhibit 1001, p. 23
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 22 of 66
`
`US 10 , 106 , 603 B2
`
`2500
`
`COL1A2
`
`Figure 17D
`
`ACTA2
`
`500
`
`2000
`
`Figure 17E
`
`Lassen - Exhibit 1001, p. 24
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`Sheet 23 of 66
`
`US 10 , 106 , 603 B2
`
`
`
`
`
`
`
`DEseq2 corrected Fold Change
`
`l wall
`
`l ' al
`
`lalal
`
`alal
`
`l
`
`
`
`wala lalalalalala
`
`Upregulated in Fibrosis
`
`
`
`. ya €
`361 WWW
`
`.
`
`wwwww
`
`* * * * *
`
`.
`
`1 X . XXX
`
`* * * * *
`
`.
`
`.
`.
`
`. . .
`1 .
`
`3
`
`Downregulated in Fibrosis
`
`Figure 18A
`
`Figure 18B
`
`Lassen - Exhibit 1001, p. 25
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 24 of 66
`
`US 10 , 106 , 603 B2
`
`TGFB1m
`Figure 18C
`
`w
`
`24h
`
`Lassen - Exhibit 1001, p. 26
`
`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 25 of 66
`
`US 10 , 106 , 603 B2
`
`
`
`Figure 18D
`
`wiiiniintindinini
`
`intiimiin niiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiin
`
`1251
`
`
`
`IL11 Expression
`
`Lassen - Exhibit 1001, p. 27
`
`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 26 of 66
`
`US 10 , 106 , 603 B2
`
`1L11 FC ( qPCR )
`
`?L11 FC ( RNA - seq )
`
`Figure 18E
`
`12111
`
`12 41
`
`OOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO
`
`
`
`rel . IL11 protein
`
`0 . 0 . 0 . 0 . 0 . . .
`
`ET - 11 PDGF TIL11
`IL13
`OSM
`No stimulus
`
`be
`
`Figure 19A
`
`Lassen - Exhibit 1001, p. 28
`
`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 27 of 66
`
`US 10 , 106 , 603 B2
`
`2000
`
`Measure ( pg / ml )
`
`V
`
`Added ( pg / ml
`
`Figure 19B
`
`( wod ) 1171
`
`16
`
`•HH . TAMAA
`
`Figure 19C
`
`Lassen - Exhibit 1001, p. 29
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 28 of 66
`
`US 10 , 106 , 603 B2
`
`Protein
`
`* *
`
`*
`
`doo
`
`Figure 19D
`
`Lassen - Exhibit 1001, p. 30
`
`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 29 of 66
`
`US 10 , 106 , 603 B2
`
`ttttttttttttttttttttttttttt
`
`. .
`
`. .
`
`.
`
`.
`
`.
`
`.
`
`* *
`
`Stimulus
`
`SMA
`
`TGFB1H
`1111
`Stimulus Collagen
`
`TGFB1
`
`Stimulus Periostin
`
`TGFB1
`
`Figure 20A
`
`Lassen - Exhibit 1001, p. 31
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 30 of 66
`
`US 10 , 106 , 603 B2
`
`
`
`Collagen Protein ( Sirius Red )
`
`Control TGFB1 1111
`
`Figure 20B
`
`IL6 ( pg / mL )
`
`W
`
`WA
`
`Control TGFB1 1L11
`
`Figure 20C
`
`Lassen - Exhibit 1001, p. 32
`
`

`

`U . S . Pater
`
`atent
`
`Oct . 23 , 2018
`
`Sheet 31 of 66
`
`US 10 , 106 , 603 B2
`
`"
`
`e n router une
`
`
`?
`
`' LL
`
`vvv
`
`AT Control TGFB1 L11 1
`
`Figure 20D
`
`MMP2 ( ng / ml )
`2 LYS
`
`Control TGFB1 L11
`
`Figure 20E
`
`Lassen - Exhibit 1001, p. 33
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 32 of 66
`
`US 10 , 106 , 603 B2
`
`
`
`Activated Fibroblasts
`
`Figure 20F
`
`Lassen - Exhibit 1001, p. 34
`
`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 33 of 66
`
`US 10 , 106 , 603 B2
`
`yo .
`HH . .
`
`OU
`
`VYYYYYYYYYYYYYYYYY
`
`w
`
`VVV
`
`w
`
`H W Merve
`
`Villalba
`
`Figure 21A
`
`1
`
`ECM ( Periostin )
`MAMA
`
`MA
`
`ECM ( Collagen )
`
`MAAVAA
`
`
`
`Activated Fibroblasts
`
`* 3 : 2 H
`Ventricular
`
`Lung
`
`Renal
`
`Fibroblast
`
`Lassen - Exhibit 1001, p. 35
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 34 of 66
`
`US 10 , 106 , 603 B2
`
`• Mouse L11 ( 100ug / kg )
`Ang | | ( 2mg / kg / day )
`o Saline - Control
`
`Ha
`
`Figure 21B
`
`w Lung
`Kidney
`
`Heart
`
`.
`
`Lassen - Exhibit 1001, p. 36
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 35 of 66
`
`| US 10 , 106 , 603 B2
`
`Liver
`buny
`Kidney
`
`LV
`
`Atrium
`
`Figure 210 .
`
`.
`
`AAAAA
`
`?? HE
`
`11 -
`
`P 0 . 0225
`
`P00268
`
`P0 , 0002
`I
`
`P00337
`
`1
`
`? ? .
`
`????
`2 P00365
`Collagen Content
`
`
`
`Lassen - Exhibit 1001, p. 37
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`Oct . 3
`
`Sheet 36 of 66
`
`US 10 , 106 , 603 B2
`
`aSMA
`
`wymiana
`
`ma soe .
`
`Figure 22A
`
`
`
`Call Proliferation
`
`wwwwwwwwww
`
`anti - 2 . 11 * * * *
`
`1
`
`Figure 22B
`
`Lassen - Exhibit 1001, p. 38
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`Sheet 37 of 66
`
`US 10 , 106 , 603 B2
`
`S .
`
`anti - tL . 19
`
`*
`-
`Figure 22C
`
`mainonnaniamising
`
`I
`
`82004 I
`
`anti - IL 11
`
`-
`Figure 22D
`
`I
`
`Lassen - Exhibit 1001, p. 39
`
`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 38 of 66
`
`US 10 , 106 , 603 B2
`
`ng / ml
`
`LLLL
`
`wwwwwwanan
`
`YYYYY
`
`Figure 22E
`
`ng / ml
`
`TGFB1
`
`Figure 22F
`
`Lassen - Exhibit 1001, p. 40
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 39 of 66
`
`US 10 , 106 , 603 B2
`
`
`
`Collagenl Protein ( Operetta )
`
`manasan
`
`
`
`
`
`Collagen Protein ( Sirius Red )
`
`ede
`
`delete
`
`+ 1 +
`
`en gure 23A
`
`* * *
`
`1674
`
`+ 11
`
`Figure 23B
`
`Lassen - Exhibit 1001, p. 41
`
`

`

`och om man sm
`
`Sheet 40 of 66
`
`US 10 , 106 , 603 B2
`
`E
`
`E
`
`U . S . Patent
`
`Oct . 23 , 2018
`
`AIRA ,
`
`LARA
`
`I
`
`
`
`
`
`
`
`?????? ??? ? ??? ???? ?? ?? ?? ?? ??? ??? ??? ??? ??? ??? ??? ??? ????
`
`
`
`
`
`111111111
`
`Figure 24
`
`wwwww . mow e donne . . . . . . . . . .
`
`mempen
`
`Lassen - Exhibit 1001, p. 42
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 41 of 66
`
`US 10 , 106 , 603 B2
`
`Myofibroblasts [ % ]
`
`M
`
`DBaseline
`OTGFB1
`
`T DANG2
`
`frentemente
`
`+ )
`IL11RA ( + + )
`Figure 25A
`
`- )
`
`
`
`Proliferating cells [ % ]
`
`C Baseline
`DTGFB1
`O ANG2
`TLT
`
`IL11RA ( + / + )
`
`( + )
`Figure 25B
`
`( m )
`
`Lassen - Exhibit 1001, p. 43
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`Sheet 42 of 66
`
`US 10 , 106 , 603 B2
`
`Collagen l ( intensity / area )
`
`ITT I
`ULEIULUULUULUU
`
`. . .
`
`C . Baseline
`DTGFB1
`
`O ANG2
`
`* 111111111111111
`
`* * *
`
`IL11RA ( + + )
`
`XXXX
`
`( + )
`Wigs
`Figure 25C
`
`I - in }
`
`C . Baseline
`DTGFB1
`DL11
`O ANG2
`
`wwwkkiinnittihakikisha hahhahahahahahaha
`
`IL11RA ( + / + )
`
`.
`
`YYYYYYYYYYYYY
`
`( - : - )
`
`( - + )
`
`Figure 250
`
`Lassen - Exhibit 1001, p. 44
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 43 of 66
`
`US 10 , 106 , 603 B2
`
`e
`
`lan
`
`ex
`
`wana
`
`w
`
`ANG2 others
`
`doen
`
`anti - L11 w
`
`W
`
`doelen
`
`online
`we
`Figure 26A
`
`com
`
`ww
`
`w it .
`
`*
`
`*
`
`Ang ! -
`L11ab
`
`Figure 26B
`
`Lassen - Exhibit 1001, p. 45
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`Sheet 44 of 66
`
`US 10 , 106 , 603 B2
`
`Collagen - HPA
`P = 0 . 019
`P 0 . 002
`
`Q Saline
`D Angil
`
`KO Saline
`Figure 27A
`Col1A2
`
`wwwwwwwwwwwwwwwww
`
`D
`
`Saline
`Angll
`
`
`
`Protein Expression
`
`wy
`
`
`
`RNA Expression
`
`tutto
`
`2
`
`CA
`ml
`
`KO Saline KO Angli
`Figure 27B
`
`Lassen - Exhibit 1001, p. 46
`
`

`

`U . S . Patent
`
`atent
`
`Oct . 23 , 2018
`
`Sheet 45 of 66
`
`US 10 , 106 , 603 B2
`
`OSMA
`
`Wh
`
`w
`
`(
`
`Saline
`Angil
`
`1
`
`2 En
`
`WT Angit
`Figure 270
`
`Fibronectin
`
`Saline
`O
`DAngli
`
`
`
`RNA Expression
`
`
`
`RNA Expression
`
`tar
`
`WT Saline WT Angli KO Saline
`
`.
`
`S
`
`Figure 27D
`
`Lassen - Exhibit 1001, p. 47
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 46 of 66
`
`US 10 , 106 , 603 B2
`
`L11RA ( - - )
`IL11RA ( + / + )
`. . . P0 . 022
`P = 0 . 005
`
`M
`
`mi
`
`2012
`
`M
`
`12 / 04
`
`-
`
`WWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWW
`
`ot
`Figure 28
`
`t
`
`Lassen - Exhibit 1001, p. 48
`
`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 47 of 66
`
`US 10 , 106 , 603 B2
`
`. . .
`
`. .
`
`* * * *
`* * * *
`
`* *
`
`*
`
`* * *
`
`2
`
`Pa '
`
`' .
`
`KO nou ( = 12 ) * * * * * * * * * *
`
`* * * * * * * * *
`
`Figure 29A
`
`Dermai fibrosis
`
`Contot muce ( 13103 * 0 *
`
`100 % * XXXXV
`
`W
`
`* *
`
`00000000000000000
`
`Figure 29B
`
`Lassen - Exhibit 1001, p. 49
`
`

`

`atent
`
`Oct . 23 , 2018
`
`Sheet 48 of 66
`
`US 10 , 106 , 603 B2
`
`Gain - of - function
`
`Y
`
`.
`
`.
`
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`Figure 290
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`Lassen - Exhibit 1001, p. 50
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`

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`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 49 of 66
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`US 10 , 106 , 603 B2
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`viwiivi
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`Figure 30A
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`# # #
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`Figure 30B
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`Lassen - Exhibit 1001, p. 51
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`

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`U . S . Patent
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`Oct . 23 , 2018
`
`Sheet 50 of 66
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`US 10 , 106 , 603 B2
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`Figure 31A
`Control
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`Figure 31B
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`Lassen - Exhibit 1001, p. 52
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`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 51 of 66
`
`US 10 , 106 , 603 B2
`
`Decoy Receptor 50aa linker
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`Myofibroblasts [ % ]
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`}
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`TGFB1 mm
`Decoy Ing / ml ] -
`
`Figure 32A
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`•6H4 : + 8
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`Decoy Receptor 33aa linker
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`117
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`1 1111111111 . 4444
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`Activ . Fibroblasts [ % ]
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`•Hit
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`TGFB1 -
`Decoy ( ng / mi )
`we
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`+ w + s 50
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`Figure 32B
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`Lassen - Exhibit 1001, p. 53
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`

`

`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 52 of 66
`
`US 10 , 106 , 603 B2
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`SEQD
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`Sequence
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`
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`Genotype Minor Allele
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`GIGTGATTGCTTAAAAAAAACTACTIC / TACATTGTTT TGAATCACACCTCACA
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`GCTCAGCTAATCAATGACCAGTCTC / C / TITTAATTCTT CTAATGCCTATATGGT
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`GTAAGOGATGTGAATCOGGTACTGA { A / GIGAAAGAG CCTGGATGCAGAGCCAGC
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`TTGATAACTTCAGCATCTGGATCACICIT ] GTGGGATT AGCATCTGTTTGTATTT
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`. . . . . . . . . . . . .
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`A
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`T
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`21
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`TTGGGGCTATAAGAGGTA
`
`Figure 33
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`GIA
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`TIC
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`GIA
`
`0 . 0248
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`07
`1 . 765
`
`FOR
`P value
`Position ( hg19 )
`SNPD
`rs 10831850
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`11 - 12566917
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`0 . 0248
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`07
`1 . 76E -
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`- - - - - - - - - - - - - - -
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`- - - - - - - - - - -
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`11 - 72599138
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`rs4756936
`
`0 . 0248
`
`0 . 0306
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`11 - 12578182 1 . 76E - 0 . 0248
`
`1 . 44
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`07
`
`2 - 223621273 2 . 70€
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`11 - 12581045
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`rs7120273
`
`- - - - - - - - - - - - - - - - - - -
`
`18895468
`
`r56485827
`
`Lassen - Exhibit 1001, p. 54
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`

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`U . S . Patent
`
`Oct . 23 , 2018
`
`Sheet 53 of 66
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`US 10 , 106 , 603 B2
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