US007345025B2
`
`(12) United States Patent
`Symonds et a1.
`
`(10) Patent N0.:
`(45) Date of Patent:
`
`US 7,345,025 B2
`Mar. 18, 2008
`
`(54) METHODS FOR GENETIC MODIFICATION
`OF HEMATOPOIETIC PROGENITOR CELLS
`AND USES OF THE MODIFIED CELLS
`
`(75) Inventors: Geo?rey P. Symonds, Rosebay (AU);
`Rafael G. Amado, Encino, CA (US);
`Lun-Quan Sun, Eastwood (AU); Janet
`L. MacPherson, Leichhardt (AU);
`Gregory C. Fanning, Bronte (AU);
`Wayne Gerlach, East Killara (AU)
`
`(73) Assignee: Johnson & Johnson Research Pty.
`Limited, Eveleigh (AU)
`
`( * ) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 0 days.
`
`(21) App1.No.: 10/192,9s0
`
`(22) Filed:
`
`Jul. 10, 2002
`
`(65)
`
`Prior Publication Data
`
`US 2004/0072771 A1
`
`Apr. 15, 2004
`
`Related US. Application Data
`
`(60) Provisional application No. 60/386,063, ?led on Jun.
`4, 2002, provisional application No. 60/343,484, ?led
`on Dec. 21, 2001, provisional application No. 60/304,
`283, ?led on Jul. 10, 2001, provisional application
`No. 60/304,127, ?led on Jul. 10, 2001.
`
`(51) Int. Cl.
`(2006.01)
`A01N 43/04
`(2006.01)
`C12Q 1/68
`(2006.01)
`C12N 15/00
`(2006.01)
`00 7H 21/02
`(2006.01)
`00 7H 21/04
`(52) us. c1. ......................... .. 514/44; 435/6; 435/911;
`435/91.31; 435/320.1; 435/455; 435/458;
`536/23.1; 536/245
`(58) Field of Classi?cation Search ................ .. 514/44;
`424/931; 435/320.1, 6, 91.1, 91.31, 455,
`435/458, 375; 536/231, 24.5
`See application ?le for complete search history.
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`3/1996 Taira et a1.
`5,500,357 A
`6/1996 McSWiggen et al.
`5,525,468 A
`5,693,535 A 12/1997 Draper et al.
`5,712,384 A
`1/1998 Syrnonds et al.
`5,911,983 A
`6/1999 Barranger et al.
`6,114,167 A *
`9/2000 Syrnonds et a1. ...... .. 435/372.3
`9/2001 Bagnis et al.
`6,287,864 B1
`5/2002 Syrnonds et al.
`2002/0058636 A1
`5/2003 Syrnonds et al.
`2003/0082158 A1
`
`FOREIGN PATENT DOCUMENTS
`
`EP
`EP
`W0
`WO
`
`0321201
`0612844
`W0 9104324
`9217211
`
`6/1989
`8/1994
`4/1991
`10/1992
`
`W0
`W0
`W0
`W0
`W0
`W0
`W0
`WO
`WO
`
`W0 9400012
`W0 9416736
`W0 9504818
`WO 95/18854
`WO 96/22368
`WO 97/47770
`WO 00/34495
`02059300
`03006612
`
`1/1994
`8/1994
`2/1995
`8/1995
`7/1996
`12/1997
`6/2000
`8/2002
`1/2003
`
`OTHER PUBLICATIONS
`
`Kohn et al. 1999 A clinical trial of retroviral-mediated transfer of a
`rev-responsive element decoy gene into CD34+ cells from the bone
`marrow of human immunode?ciency virus-l-infected children
`Blood, vol. 94 pp. 368-371.*
`Gervaix et al. 1997 Gene therapy targeting peripheral blood cd34+
`hematopoietic stem cells of HIV-infected individuals Human Gene
`Therapy 8:2229-2238.*
`Lee et al. 2002 Expression of small interfering RNAs targeted
`against HIV-1 rev transcripts in human cells. Nature Biotechnology
`19:500-505.*
`Slobod et al. 1996 Mobilization of CD34+ progenitor cells by
`granulocyte colony-stimulating factor in human immunode?ciency
`virus type l-infected individuals. Blood vol. 99(9):3329-3335.*
`U.S. Appl. No. 08/375,291, ?led Jan. 18, 1995, application With
`pending claims as of Jan. 10, 2002 (Exhibit 6).
`Amado et al., “Effects of Megakaryocyte Growth and Development
`Factor on Survival and Retroviral Transduction of T Lymphoid
`Progenitor Cells”, Human Gene Therapy 9: 173-183 (1998) (Exhibit
`8).
`Barinaga, Science 262:1512 (1993) (Exhibit 9).
`BerZal-HerranZ, A. et al. “Essential Nucleotide Sequences and
`Secondary Structure Elements of the Hairpin RiboZyme.” EMBO
`12:2567-2574 (1993) (Exhibit 10).
`Bevec, D. et al., “Inhibition of Human Immunode?ciency Virus
`Type 1 Replication in Human T Cells by Retroviral-Mediated Gene
`Transfer of a Dominant-Negative Rev Trans-Activator.” Proc. Natl.
`Acad. Sci. 89:9870-9874 (1992) (Exhibit 11).
`Bischofberger, N. and Wagner, R.W. “Antisense Approaches to
`Antiviral Agents”. Virology 3:57-66 (1992) (Exhibit 12).
`Buckley, Abstract #446, [online] 2001 Retrieved from Internet:
`<URL:http://WWW4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 13).
`Cech et al., RiboZyme engineering Curr. Opin. Structural Biol.
`21605-609 (Exhibit 14).
`Chen, C-J. et al., “Inhibition of HIV-1 Replication by Novel
`Multitarget Ribozymes”. Ann. NY. Acad. Sci. 660:271-273 (1992)
`(Exhibit 15).
`
`(Continued)
`Primary Examinerilane Zara
`(74) Attorney, Agent, or Firmilohn P. White; Cooper &
`Dunham LLP
`
`(57)
`
`ABSTRACT
`
`Described are compositions and methods relating to gene
`therapy, particularly as applied to hematopoietic progenitor
`(HP) cells, to transduced cells and methods of obtaining
`them, and to methods of using them to provide prolonged
`engraftment of modi?ed hematopoietic cells in human sub
`jects. The invention particularly relates to ex vivo gene
`therapy of HP cells for treatment or prevention of HIV
`infection.
`
`18 Claims, 21 Drawing Sheets
`
`Benitec - Exhibit 1015 - page 1
`
`

`
`US 7,345,025 B2
`Page 2
`
`OTHER PUBLICATIONS
`
`Chen, C-J. et al., “Multitarget-Ribozyme directed to Cleave at up to
`Nine Highly Conserved HIV-1 env RNA Regions Inhibits HIV-1
`Replication-Potential Effectiveness Against Most Presently
`Sequenced HIV-1 Isolates”. Nucleic Acids Res. 2014581-4589
`(1992) (Exhibit 16).
`ChoWrira, B.M. “Four Ribose 2‘-Hydroxyl Groups Essential for
`Catalytic Function of the Hairpin RiboZyme”. J. Biol. Chem.
`268119458-19462 (1993) (Exhibit 17).
`Chu, “Retrovirus-mediated gene transfer into human hematopoietic
`stem cells”, J. Mol. Med. 761184-192 (1998) (Exhibit 18).
`Cournoyer, D. and Caskey, C.T. “Gene Therapy of the Immune
`System” Annu. Rev. Immunol. 111297-329 (1993) (Exhibit 19).
`Crisell, P. et al., “Inhibition of HIV-1 Replication by RiboZymes that
`Show Poor Activity in vitro” Nucleic Acids Res. 2115251-5255
`(1993) (Exhibit 20).
`Donahue et al., J. Vir. 621722 (1988) (Exhibit 21).
`Dropulic, B. et al., (1992) J. Virol. 6611432-1441 (Exhibit 22).
`Dropulic, B. et al., “Ribozymes: Use As Anti-HIV Therapeutic
`Molecules”. Antisense Res. Dev. 3187-94 (1993) (Exhibit 23).
`Gervaix, “Gene Therapy Targeting Peripheral Blood CD34+ Gene
`Hematopoietic Stem Cells of HIV-Infected Individuals” Human
`Therapy 812229-2238 (1997) (Exhibit 24).
`Goodchild et al., “Antisense Antivirals”. Antisense Res. Dev. 1:361
`364 (1991) (Exhibit 25).
`Goodchild et al., Enhancement of riboZyme catalytic activity, by a
`contigous oligodeoxynucleotide (facilitator) and by 2‘-O-methyla
`tion Nucleic Acids Res. 2014607-4612 (1992) (Exhibit 26).
`Halene, “Gene Therapy Using Hematopoietic Stem Cells: Sisyphus
`Approaches the Crest”, Human Gene Therapy 1111259-1267 (2000)
`(Exhibit 27).
`Han et al. PNAS 8814313 (1991) (Exhibit 28).
`Heidenreich, O. and Eckstein F. “Hammerhead RiboZyme-Mediated
`Cleavage of Long Terminal Repeat RNA of Human
`Immunde?ciency Virus Type 1”. J. Biol. Chem. 26711904-1909
`(1992) (Exhibit 29).
`Homann, M. et al., “Incorporation of the Catalytic Domain of a
`Hammerhead RiboZyme Into Antisense RNA Enhances Its Inhibi
`tory Effect on the Replication of Human Immunode?ciency Virus
`Type 1”. Nucleic Acids Res. 2112809-2814 (1993) (Exhibit 30).
`Johnston et al. “Present Status and Future Prospects for HIV
`Therapies”, Science 26011286-1293 (1993) (Exhibit 31).
`Joseph, S. et al. “Optimization of an Anti-Hairpin RiboZyme by in
`Vitro Selection” J. Biol. Chem. 268124515-24518 (1993) (Exhibit
`32).
`Joseph, S. et al. “Substrate Selection Rules for the Hairpin
`RiboZyme Determined by in Vitro Selection, Mutation, and Analy
`sis of Mismatched Substrates”. Genes and Development 71130-138
`(1993) (Exhibit 33).
`Lever et al. J. Vir. 6314085 (1989) (Exhibit 34).
`Levy. J. “Pathogenesis of human Immunode?ency Virus Infection”.
`Microbiol. Rev. 571183-289 (1993) (Exhibit 35).
`LisZieWicZ, J. et al., “Inhibition of Human Immunode?ciency Virus
`Type 1 Replication By Regulated Expression of a Polymeric Tat
`Activation Response RNA Decoy as a Strategy For Gene Therapy
`in AIDS”. Proc. Natl. Acad. Sci. 9018000-8004 (1993) (Exhibit 36).
`Liu, JM, Abstract #78, [online] 1994 Retrieved from Internet:
`<URL1http1//WWW4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 37).
`Lo, K.M. et al., “Inhibition of Replication of HIV-1 by Retroviral
`Vectors Expressing tat-Antisense and Anti-tat RiboZyme RNA”.
`Virology 1901176-183 (1992) (Exhibit 38).
`Lori F. et al., “Rapid Protection Against Human High HIV-1
`Immunode?ciency Virus Type 1 (HIV-1) Replication Mediated by
`E?‘iciency Non-retroviral Delivery of Genes Interfering With tat and
`gag”. Gene Therapy 1127-31 (1994) (Exhibit 39).
`Mann et al. J. Vir. 541401 (1985) (Exhibit 40).
`Mountain, “Gene therapy: the ?rst decade”, Tibtech vol. 18 pp.
`119-128 (2000) (Exhibit 41).
`Ohkawa, J. et al., “Importance of Independence in RiboZyme
`Reactions: Kinetic Behavior of Trimmed and of Simply Connected
`
`Multiple RiboZymes Wit Potential Activity Against Human
`Immunode?ciency Virus”. Proc. Natl. Acad. Sci. 90111302-11306
`(1993) (Exhibit 42).
`Ohkawa, J. et al., “Multiple Site-Speci?c Cleavage of HIV RNA by
`Transcribed RiboZymes from Shotgun-Type Trimming Plasmid”.
`Nucleic Acids Symp. Ser. 291121-122 (1993) (Exhibit 43).
`Ojwang, J .O. et al., “Inhibition of Human Immunode?ciency Virus
`Type 1 Expression by a Hairpin RiboZyme”. Proc. Natl. Acad. Sci.
`89110802-10806 (1992) (Exhibit 44).
`Olmsted et al. PNAS 8618088 (1989) (Exhibit 45).
`Ratner et al. Nature 3131277 (1985) (Exhibit 46).
`Romano, “Latest Developments in Gene Transfer Technology:
`Achievements, Perspectives, and Controversies over Therapeutic
`Applications”, Stem Cells 18:19-39 (2000) (Exhibit 47).
`Rossi, JJ et al., “Catalytic Antisense RNA (Ribozymes): Their
`Potential and Use as Anti-HIV-l Therapeutic Agents”. Innov. In
`Antiviral Dev. & the Detecting of Virus Infection 312195-109
`(1992) (Exhibit 48).
`Rossi, JJ et al., “RiboZymes as Anti-HIV-l Therapeutic Agents:
`Principles, Applications, and Problems”. AIDS Res. Hum.
`Retroviruses 81183-189 (1992) (Exhibit 49).
`Rossi, JJ et al., “RiboZymes as Therapies for AIDS”. Ann. NY.
`Acad. Sci. 6161184-200 (1990) (Exhibit 50).
`Sarver, N. “Ribozymes: A New Frontier in Anti-HIV Strategy”.
`Antisense Res. Dev. 11373-378 (1991) (Exhibit 51).
`Sarver, N. “RiboZymes as Potential Anti-HIV-l Therapeutic
`Agents”. Science 24711222-1225 (1990) (Exhibit 52).
`Sarver et al., Aids Res. Rev. 21259-285 (1992) (Exhibit 53).
`Shimayama T. et al., “Cleavage of the Highly Conserved Symp.
`Hairpin-Loop Region of HIV-1 by Synthetic RiboZymes”. Nucleic
`Acids Sr. 291177-178 (1993) (Exhibit 54).
`Stull et al. Pharm. Res. 121465 (1995) (Exhibit 55).
`Taylor, NR. and Rossi J.J., “RiboZyme-Mediated Cleavage of a
`HIV-1 gag RNA: The Effects of Nontargeted Sequences and Sec
`ondary Structure on RiboZyme Cleavage Activity”, Antisense Res.
`Dev. 11173-186 (1991) (Exhibit 56).
`Thill, G. et al., “Structural and Sequence Elements Required for the
`Self-Cleaving Activity of the Hepatitis Delta Virus RiboZyme” ,
`Biochemistry 3214254-4262 (1993) (Exhibit 57).
`Weerasinghe, M. et al., “Resistance to Human Immunode?ciency
`Virus Type 1 (HIV-1) Infection in Human CD4+ Lymphocyte
`Derived Cell Lines Conferred by Using Retroviral Vectors Express
`ing an HIV-1 RNA-Speci?c RiboZyme”, J. Virol 6515531-5534
`(1991) (Exhibit 58).
`Yamada, O. et al., “Intracellular Immunization of Human T Cells
`With a Hairpin RiboZyme Against Human Immunode?ciency Virus
`Type 1”, Gene Therapy 1138-45 (1994) (Exhibit 59).
`Yu, M. et al., “A Hairpin RiboZyme Inhibits Expression of Diverse
`Strains of Human Immunode?ciency Virus Type 1”, Proc. Natl.
`Acad. Sci. 9016340-6344 (1993) (Exhibit 60).
`Zaia, J.A. et al., “Status of RiboZyme and Antisense-Based Devel
`opmental Approaches for Anti-HIV-l Therapy”, Ann. NY. Acad.
`Sci 660195-106 (1992) (Exhibit 61).
`Abonour, Abstract #172, [online] 1997 Retrieved from Internet:
`<URL1http1//WWW4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 1).
`Abonour, “E?‘icient retrovirus-mediated transfer of the multidrug
`resistance 1 gene into autologous human lone-term repopulating
`hematopoietic stem cells”, Nature Medicine, vol. 6, No. 6, pp.
`652-658 (2000) (Exhibit 2).
`Amado, Systemix #261, [online] 1998 pp. 11-13 <URL1http1//
`WWW4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 3).
`Amado, Systemix #277, [online] 1998 pp. 15-16 <URL1http1//
`WWW4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 4).
`Amado et al., “A Phase I Trial of Autologous CD34+ Hematopoietic
`Progenitor Cells Transduced With an Anti-HIV RiboZyme”, Human
`Gene Therapy 1012255-2270 (1999) (Exhibit 5).
`Barranger, JA, Scienti?c Summary #46, [online] 1993 <URL1http1//
`WWW4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 6).
`Bauer, Scienti?c Abstract #204, [online] 1997 <URL1http1//WWW4.
`od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 7).
`Bordignon et al., “Gene Therapy in Peripheral Blood Lymphocytes
`and bone Marrow for ADA’ Immunode?cient Patients” Science vol.
`270, 470-475 (1995) (Exhibit 8).
`
`Benitec - Exhibit 1015 - page 2
`
`

`
`US 7,345,025 B2
`Page 3
`
`Carabasi, Systemix #340 (Protocol 108), [online] 1999 pp. 13-16
`<URL:http://www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 9).
`CavaZZana-Calvo, “Gene Therapy of Human Severe Combined
`Immunode?ciency (SCID)-X1 Disease”, Science vol. 288, 669-672
`(2000) (Exhibit 10).
`Champlin, Scienti?c Abstract #44, [online] 1993 <URL:http://
`www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 11).
`Conant, EnZo Therapeutics, Inc. Scienti?c Abstract #230, [online]
`1998 <URL:http://www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit
`12).
`Cowan, Abstract #143, [online] 1996 <URL:http://www4.od.nih.
`gov/oba/rac/hgtprep.asp> (Exhibit 13).
`Croop, Scienti?c Abstract #370, [online] 2000 <URL:http://www4.
`od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 14).
`Dinauer, Scienti?c Abstract # 196, [online] 1997 <URL:http://
`www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 15).
`Douer, Scienti?c Abstract #92, [online] 1994 pp. 174-175
`<URL:http://www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 16).
`Dunbar et al., Abstract # 47, [online] 1993 pp. 174-175 <URL:http://
`www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 17).
`Dunbar et al., “Retrovirally Marked CD34’Enriched Peripheral
`Blood and Bone Marrow Cells Contribute to Long-Term Engraft
`ment After Autologous Transplantation”, Blood, vol. 85, No. 11, pp.
`3048-3057 (1995) (Exhibit 18).
`Gabarre et al., “High-dose therapy and autologous haematopoietic
`stem-cell transplantation for HIV-1-associated lymphoma”, The
`Lancet, The Lancet, vol. 355, pp. 1071-1072 (2000) (Exhibit 19).
`Holodinity, Systemix Abstract #333, [online] 1999 pp. 13-16
`<URL:http://www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 20).
`Kohn et al., “Engraftment of gene-modi?ed umbilical cord blood
`cells in neonates with adenosine deaminase de?ciency”, Nature
`Medicine, vol. 1, No. 10, pp. 1017-1023 (1995) (Exhibit 21).
`Kohn, Scienti?c Abstract # 147, [online] 1996 <URL:http://www4.
`od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 22).
`Kohn/Zaia, Scienti?c Abstract # 153, [online] 1996 <URL:http://
`www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 23).
`Kohn et al., “T lymphocytes with a normal ADA gene accumulate
`after transplantation of transduced autologous umbilical cord blood
`CD34+ cells in ADA-de?cient SCID neonates”, Nature Medicine
`vol. 4: 775-780 (1998) (Exhibit 24).
`Kohn et al., “A Clinical Trial of Retroviral-Mediated Transfer of a
`rev-Responsive Element Decoy Gene Into CD34+ Cells From the
`Bone Marrow of Human Immunode?ciency Virus-l-Infected Chil
`dren”, Blood, vol. 94, No. 1, pp. 368-371 (1999) (Exhibit 25).
`Kohn et al. Scienti?c Abstract #390, [online] (Mar. 2000)
`<URL:http://www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 26).
`Laurence, J ., EnZo Therapies #443, [online] 2001, pp. 1-3
`<URL:http://www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 27).
`Law et al., “Mobilization of peripheral blood progenitor cells for
`human immunode?ciency virus-infected individuals”, Experimen
`tal Hematology 27:147-154 (1999) (Exhibit 28).
`Liu, “Regulated Expression of a Dominant Negative Form of Rev
`Improves Resistance to HIV Replication in T Cells”. Gene Therapy
`1:32-37 (1994) (Exhibit 29).
`Malech, Scienti?c Abstract #104, [online] 1995 <URL:http://
`www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 30).
`Malech, Scienti?c Abstract #231, [online] 1998 <URL:http://
`www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 31).
`Michallet et al., “Transplantation with selected autologous periph
`eral blood CD34+Thyl+ hematopoietic stem cells (HSCs) in multiple
`myeloma: Impact of HSC dose on engraftment, safety, and immune
`reconstitution”, Experimental Hematology 28:858-870 (2000)
`(Exhibit 32).
`Novelli et al., 2000 (Abstract #3448) A Phase I Trial of Retroviral
`Mediated Gene Therapy of Gaucher Disease, Clinically-Related
`Gene Therapy Studies 798a (Exhibit 33).
`O’Shaughnessy, Abstract #54, [online] 1993 <URL:http://www4.
`od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 34).
`Sandhaus et al., “A simpli?ed method of CD34+ cell determination
`for peripheral blood progenitor cell transplantation and correlation
`with clinical engraftment”, Experimental Hematology 26:73-78
`(1998) (Exhibit 35).
`
`Schuening, Scienti?c Abstract #61, [online] 1993 <URL:http://
`www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 36).
`Buckley, Scienti?c Abstract # 446, [online] 2001 <URL:http://
`www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 37).
`Tisdale, Precis Abstract # 341, [online] 1999 <URL:http://www4.
`od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 38).
`Walsh, Scienti?c Abstract #291, [online] 1999 <URL:http://www4.
`od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 39).
`Krishnan et al- Non-technical Abstract #218, [online] 1997
`<URL:http://www4.od.nih.gov/oba/rac/hgtprep.asp> (Exhibit 40).
`Lee, Benhur, “An Intricate Web: Chemokine Receptors, HIV-1 and
`Hematopoiesis”, Stem Cells 1998; 16:79-88 (Exhibit 41).
`de Feyter, Rob, Technology evaluation: HIV riboZyme gene therapy,
`Gene Shears Pty Ltd, Current Opinion in Molecular Therapeutics,
`2000, vol. 2, No. 3, pp. 332-335 (Exhibit 43).
`Calenda et al., “The Effects of HIV on Hematopoiesis” European
`Journal of Haematology, 1992, vol. 48, pp. 181-186 (Exhibit 44).
`Morel et al., “Hematologic Recovery in Mice Transplanted with
`Bone
`Marrow
`Stem
`Cells
`Expressing
`Anti-Human
`Immunode?ciency Virus Genes” Human Gene Therapy, Nov. 20,
`1999, vol. 10 pp. 2779-2787 (Exhibit 45).
`Mautino et al., “Gene Therapy of HIV-1 Infection Using Care
`Lentiviral Vectors Expressing Anti-HIV Genes”, AIDS Patient and
`STDs, 2002, vol. 16, No. 1, pp. 11-26 (Exhibit 46).
`Bauer et al., “Gene Therapy for Pediatric AIDS”, Annals of the New
`York Academy of Sciences, 2000, vol. 918, pp. 318-329 (Exhibit
`47).
`Sadelain et al., “Issues in the Manufacture and Transplantation of
`Genetically Modi?ed Hematopoietic Stem Cells” Current Opinion
`in Hematology, 2000, vol. 7, pp. 364-377 (Exhibit 48).
`Novina et al., “siRNA-directed inhibition of HIV-1 infection”,
`Nature Medicine, Jul. 2002, vol. 7, No. 8 pp. 681-686 (Exhibit 50).
`Poeschla et al., “Developement of HIV Vectors for Anti-HIV Gene
`Therapy”, Proceedings of the National Academy of Sciences, Oct.
`1996, vol. 93, pp. 11395-11399 (Exhibit 52).
`Ijichi et al., “In vivo induction of human immunode?ciency to virus
`type 1 entry into nucleus-free cells by CD4 gene transfer
`hematopoietic stem cells: a hypothetical possible strategy for thera
`peutic intervention”, Medical Hypothesis, 2002, vol. 59, No. 1, pp.
`24-34 (Exhibit 53).
`Lawrence, D., “RNAi Could Hold Promise in the Treatment of
`HIV”, The Lancet, Jun. 8, 2002, vol. 359, p. 2007 (Exhibit 54).
`Declaration ofGeolfrey P Symonds, Ph.D. Under 37 CPR. § 1.132
`?led on May 9, 2000 with the United States Patent and Trademark
`Of?ce in connection with US. Appl. No. 08/375,291, pp. 1-3,
`including Exhibits 1-2, copies of references as Exhibits 1-11 and
`Exhibits B-G.
`Declaration ofGeolfrey P Symonds, Ph.D. Under 37 CPR. § 1.132
`?led on Nov. 28, 2001 with the United States Patent and Trademark
`Of?ce in connection with US. Appl. No. 08/375,291, pp. 1-3,
`including Exhibit 1.
`Israeli Application No. 88683/2 (Abstract only).
`Israeli Application No. 98543/2 (Abstract only).
`Akhtar et al., (1996) “Anti-HIV Therapy with Antisense
`Oligonucleotides and RiboZymes: Realistic Approaches or Expen
`sive Myths”, J. Antimicrob. Chemother, vol. 38, pp. 159-165.
`Amado et al. “A Phase I gene therapy clinical study of Autologous
`CD34+ cells transduced with an anti-HIV-l riboZyme”, 6th Confer
`ence on Retrovirus and Opportunistic Infections, Chicago, 1999.
`Amado et al. “Development of genetically protected T-lymphocytes
`from transduced
`hematopoietic
`progenitors
`in human
`immunode?ciency virus-1 infected patients” 14th International
`AIDS Conference, Barcelona, 2002.
`Amado et al. “Multilineage engraftment and preferential survival of
`riboZyme containing CD4+ T-lymphocytes derived from CD34+
`progenitor cells in a phase I study in HIV-1”, 5Lh Am Soc Gener
`Therapy Meeting, 2002.
`Carr et al. “A Phase I Gene Therapy Study showing safety, feasi
`bility and sustained survival of anti-HIV-I transduced CD4+ T
`lymphocytes”, 6th Conference on Retroviruses and Opportunistic
`Infections, Chicago, 1999.
`Christolfersen et al. (1995) “RiboZymes as Human Therapeutic
`Agents”, J. ofMed. Chemistry, vol. 38, pp. 2023-2037.
`
`Benitec - Exhibit 1015 - page 3
`
`

`
`US 7,345,025 B2
`Page 4
`
`Cooper et al. (1999) “A marker study of therapeutically transduced
`CD4+ peripheral blood lymphocytes in HIV discordant twins”,
`Human Gene Therapy, vol. 10, pp. 1401-1421.
`Ely et al. “Retrovirus vectors and gene therapy for HIV”, Invited
`Speaker, Annual Meeting of the Australian Society for Microbiol
`ogy, Hobart, 1998.
`Ely et al. (1998) “Anti-HIV ribozymes in the inhibition of HIV and
`AIDS”, Invited Review Biogenic Amines.‘ Special Issue on Gene
`Therapy, Ed. R Bertolotti, vol. 1, pp. 113-135.
`Ely et al. “Conducting gene therapy clinical trials-paving a new
`path in clinical research”, International Clinical Trials Symposium.‘
`Improving Health in the New Millennium, Sydney, 1999.
`Fanning et al. (2000) “Anti HIV ribozyrnes in the inhibition of HIV
`and AIDS”, Invited Review Progress in Gene Therapy.‘ Basic and
`Clinical Frontiers, Eds. R Bertolotti, SH Parvez and T Nagatsu,
`VSP Utrecht, pp. 433-454.
`Fanning et al. (2002) “Ribozymes as gene therapeutic agents for
`HIV/AIDS; a potential paradigm shift”, Invited Review Pathogenic
`Genomics.‘ Impact on Human Health, Ed. KJ Shaw, Humana Press
`Inc, Totowa, NJ, pp. 39-51.
`Gerlach et al. (1989) “Synthetic Ribozymes for in vivo inactivation
`of prokaryotic or eukaryotic RNA transcripts”, Chemical Abstracts,
`vol. 112, pp. 51284.
`Gerlach et al. (1996) “Ribozymes in HIV gene therapy”, Organisa
`tion for Economic Cooperation and Development, Gene Delivery
`Systems, A state-of-the-Art Review, pp. 303-307.
`Harrison and Lever (1992) “The Human Immunode?ciency Virus
`Type 1 Packaging Signal and Major Splice Donor Region Have A
`Conserved Stable Secondary Structure”, .I. of I/lrology, vol. 66, pp.
`4144-4153.
`Haselo?C & Gerlach (1988) “Simple RNA Enzymes with New and
`Highly Speci?c Endoribonuclease Activities”, Nature, vol. 334, pp.
`585-591.
`“Therapy for Human
`(1993),
`D’Aquila
`and
`Hirsch
`Immunode?ciency Virus Infection”, New England .I. of Med., vol.
`328, pp. 1686-1695.
`Jennings et al. (1994) “DNA-Armed Ribozymes and Minizymes”,
`Chemical Abstracts, vol. 121, p. 275264b.
`Jennings et al. (1991) “Minizymes: Oligonucleotide Endonucleases
`and their use in Disease Treatment”, Chemical Abstracts, vol. 117,
`p. 127376s.
`Knop et al. (1998) “Arti?cial capillary culture: expansion and
`retroviral transduction of CD4+T- lymphocytes for clinical appli
`cation”, Gene Therapy, vol. 6, pp. 373-384.
`Leavitt et al. (1994) Transfer of an anti-HIV-1 ribozyrne gene into
`primary human lymphocytes, Human Gene Therapy, vol. 5, pp.
`11 15-1 120.
`Levy, (1994) “Antiviral Approaches”, HIV and the Pathogenesis of
`AIDS, ASM Press, pp. 217-236.
`Lowenstein et al. (1997) “Inhibition of Moloney murine leukemia
`virus (MoMLV) by a retroviral vector LNL6, carrying ribozyrnes,
`targeted to the 5‘ non-coding sequence”, J. Gen. I/lrol, vol. 78, pp.
`2587-2590.
`Macpherson et al.(1999) “Ribozymes in gene therapy of HIV-1”,
`Invited Review, Frontiers in Bioscience, vol. 4, pp. 497-505.
`Ngok, (2004) Clinical gene therapy research utilizing ribozyrnes:
`application to the treatment of HIV/AIDS, Methods Mol. Biol., vol.
`252, pp. 581-598.
`Raponi et al.(1999) “Gene delivery technology: non-viral and viral
`vector systems”, Invited Review Ex I/lvo Cell Therapy, Eds.
`Schindheilm and Nordon, Academic Press, Chapter 14, pp. 293
`322.
`Richman, (1990) “The Clinical Use of Anti-HIV Agents,” Design of
`Anti-AIDS Drugs, ed. E. de Clercq, Pharmacochemistry Library,
`Elsevier, vol. 4, pp. 339-366.
`Rigden et al. (1999) “ The use of ribozyme gene therapy for the
`inhibition of HIV replication and its pathogenic sequelae”, Invited
`Review Intracellular Ribozyme Applications.‘ Principles and Pro
`tocols, Eds. Couture and Rossi, pp. 255-270.
`Rigden et al. (2000) “The use of ribozyme gene therapy for the
`inhibition of HIV replication and its pathogenic sequelae”, Invited
`Review Current Issues in Molecular Biology, vol. 2, pp. 61-69.
`
`Sioud, M. (1994) “Tumor Necrosis Factor-Alpha Ribozyrnes and
`degradation-resistant RNA derivatives linked to TNF-alpha
`Ribozymes”, Chemical Abstracts, vol. 121, p. 127034m.
`Smythe et al. “Gene therapy: antisense and ribozyme strategies for
`HIV-1 infection”, Lorne Genome Conference, 1995.
`Sun, et al. (1994) “Ribozyme-Mediated Suppression of Moloney
`Murine Leukemia Virus and Human Immunode?ciency Virus Type
`I Replication in Permissive Cell Lines” PNAS, vol. 91, pp. 9715
`9719.
`Sun et al. “Ribozyme and antisense constructs confer resistance to
`HIV-1 infection in hemopoietic cells”, Lorne Genome Conference,
`1995.
`Sun, et al. (1995) “Target Sequence-Speci?c Inhibition of HIV-1
`Replication by Ribozymes Directed to tat RNA”, Nucleic Acid Res. ,
`vol. 23, pp. 2909-2913.
`Sun et al. (1995) “Ribozyme and antisense constructs confer resis
`tance to HIV-1 infection in hemopoietic cells”, Ribozymes: Basic
`Science and Therapeutic Applications, Keystone Meeting, Abstract:
`J. Cell. Biochem. Supp 19A, p. 223.
`Sun et al.(1995) “Resistance to human immunode?ciency virus type
`1 infection conferred by transduction of human peripheral blood
`lymphocytes with ribozyme, antisense or polymeric TAR con
`structs”, Proc. Natl. Acad Sci. USA, vol. 92, pp. 7272-7276.
`Sun et al. (1995) “Antisense and ribozyme strategies for HIV-1
`infection”, Invited Review Drugs New Perspect., vol. 8, pp. 325
`331.
`Sun et al. “The use of ribozyme constructs for a gene therapy
`approach to AIDS”, Invited Plenary Speaker, T he 1996 Palm Springs
`Symposium on HI V/AIDS, Palm Springs, California, 1996, Brochure
`only.
`Sun et al. “Ribozyme constructs in a gene therapy approach for
`AIDS”, Invited Speaker, Gene Therapy of Cancer, AIDS and
`Genetic Disorders, Trieste, Italy, 1996.
`Sun et al. “The use of ribozyme constructs for a gene therapy
`approach to AIDS”, XIth International Conference on AIDS,
`Vancouver, Canada, 1996.
`Sun et al. (1997) “Anti-HIV ribozymes”, Invited Review. Molecular
`Biotechnology, vol. 7, pp. 241-251.
`Sun et al. (1998) “The design, production and validation of an
`anti-HIV type 1 ribozyme”, (Invited Review) Methods in Molecular
`Medicine, Therapeutic Applications of Ribozyrnes, Ed. KJ Scanlon,
`Humana Press USA, pp. 51-64.
`Sun et al. (1998) “Retroviral delivery of ribozymes”, Invited
`Review Ribozymes in the Gene Therapy of Cancer, Eds. Scanlon,
`K] and Kashani-Sabet, M, pp. 87-100.
`Sun et al. (1999) “Exogenous gene transfer into lymphoid and
`haematopoietic progenitor cells”, Invited Review Ex I/lvo Cell
`Therapy, Eds. Schindheilm, K and Nordon, R, Academic Press,
`Chapter 9, pp. 179-195.
`Sun et al. (1999) “Gene Therapy in haematopoietic cells”, Cancer
`Forum vol. 23, pp. 2-5.
`Symonds et al. “Gene therapeutic approaches to AIDS”, Biotech '94
`Biotechnology Against AIDS,‘ From Basic Science to Prevention,
`Diagnosis and Therapy.
`Symonds et al. “Gene therapy for AIDS: phase I clinical trials”,
`Meeting of the Aust Soc HIV Medicine, 1997.
`Symonds, G., “Gene therapy for HIV: Issues and implications of the
`approach”, Invited Speaker, Clinical Trials Strategy Conference,
`Sydney 2000.
`Symonds, G., “Trials and tribulations of the gene transfer process”,
`Invited Speaker AusBiotech National Biotechnology & Investment
`Forum, Melbourne, 2002.
`Symonds, G., “Inhibition of HIV replication by the use of ribozyrnes
`targeted to the genome of HIV”, Invited Speaker Australasian
`Society of HI V Medicine Conference, Sydney, 2002.
`Uhlmann (1990) “Antisense Oligonucleotides: A New Therapeutic
`Principle”, Chemical Reviews, vol. 90, pp. 544-584.
`Verma et al. (1997) “Gene Therapy Promises, problems, and Pros
`pects”, Nature, vol. 389, pp. 239-242.
`Wang et al. (1998) “Preclinical Characterization of an Anti tat
`Ribozyme for Therapeutic Application”, Human Gene Therapy, vol.
`9, pp. 1283-1291.
`
`Benitec - Exhibit 1015 - page 4
`
`

`
`US 7,345,025 B2
`Page 5
`
`Yu et al. (1994) “Progress towards gene therapy for HIV infection”,
`Gene Therapy, vol. 1, pp. 12-26.
`Bahner, et al. “Transduction of Human CD34+ Hematopoietic
`Progenitor Cells by a Retroviral Vector Expressing an RRE Decoy
`Inhibits Human Immunode?ciency Virus Type 1 Replication in
`Myelomonocytic Cells Produced in Long-Term Culture,” J. I/lrol.
`7014352-4360.
`Deonarain 1998, Expert Opin Ther Pat, vol. 8, pp. 53-69.
`Bauer et al., (1997) “Inhibition of Human Immunode?ciency
`Virus-1 (HIV-1) Replication After Transduction of Granulocyte
`Colony-Stimulating Factor-Mobilized CD 34* Cells From HIV-1
`Infected Donors Using Retroviral Vectors Containing Anti-HIV-1
`Genes,” Blood 89 12259-2267.
`Bodine et al., (1998) “Improved Amphotropic Retrovirus-Mediated
`Gene Transfer into Hematopoietic Stem Cells,” Ann N Y Acad Sci
`8501139-50.
`Bonyhadi et al., (1997) “RevM10-Expressing T Cells Derived In
`Vivo from Transduced Human Hematopoietic Stem-Progenitor
`Cells Inhibit Human Immunode?ciency Virus Replication,” J Wrol
`7114704-16.
`Briones et al., (1999) “Retroviral gene transfer into human
`hematopoietic cells an in vitro kinetic study,” Haematologica 84
`1483-488.
`Case et al., (1999) “Stable transduction of quiescent CD34+CD38’
`Human hematopietic cells by HIV-1-based lentiviral vectors,” Proc
`Natl Acad Sci USA 9612988-93.
`Cavazzana-Calvo et al., (2001) “Gene therapy of severe combined
`immunode?ciencies,” J. Gene Med. 31201-201.
`Chatterjee et al., (2001) “Transduction of Primitive Human marrow
`and Cord Blood-Derived Hematopoietic Progenitor Cells with
`Adeno-Associated Virus Vectors,” Blood 931 1882-1894.
`Cohen-Haguenauer et al., (1999) “E?‘icient Transduction of
`Hemopoietric CD34+ Progenitors of Human Origin Using an Origi
`nal Retroviral Vector Derived from Fr-MuLV-FB29: In Vitro
`Assessment,” Hum Gene Ther 91207-16.
`Cui et al., (2002) “Targeting transgene expression to antigen
`presenting cells derived from lentivirus-transduced engrafting
`human hematopoietic stem/progenitor cells,” Blood 99 1399-408.
`Dao et al., (1997) “FL T3 Ligand Preserves the Ability of Human
`CD34+ Progenitors to Sustain Long-Term Hematopoiesis in
`Immune-De?cient Mice After Ex Vivo Retroviral-Mediated
`Transduction,” Blood 89 1446-456.
`Dao et al., (1998) “Reduction in levels of the cyclin-dependent
`kinase inhibitor p27kip'l coupled with transforming growth factor [5
`neutralization induces cell-cycle entry and increases retroviral
`transduction of primitive human hematopoietic cells,” Proc Natl
`Acad Sci USA 95113006-11.
`Dao et al., (1998) “Adhesion to Fibronectin Maintains Regenerative
`Capacity During Ex Vivo Culture and Transduction of Human
`Hematopoietic Stem and Progenitor Cells,” Blood 9214612-21.
`Dunbar et al., (1996) “Retroviral Mediated Transfer of the cDNA for
`Human Glucocerebrosidase into Hematopoietic Stem Cells of
`Patients with Gaucher Disease. A Phase I Study,” Human Gene T her
`71231-253.
`of the
`Transfer
`“Retroviral
`(1998)
`al.,
`Dunbar et
`Glucocerobrosidase Gene into CD34+ Cells from Patients with
`Gaucher Disease: In Vivo Detection of Transduced Cells without
`Myeloablation,” Human Gene Ther 912629-2640.
`Eglitis and Schneiderman, (1997), “Transduction of Human
`Hematopoietic Progenitor Cells with Retroviral Vectors Based on
`the Gibbon Ape Leukemia Virus,” Biochem Biophys Res Commun
`2311477-80.
`Fan et al., (2000) “E?‘icient Adenoviral Vector Transduction of
`Human Hematopoietic SCID-Repopulating and Long-Term Cul
`ture-Initiating Cells,” Hum Gene Ther 1111313-27.
`Fehse et al., (1997) “Selective Immunoaf?nity-Based Enrichment of
`CD34+