Top

Published in:

01-02-2018 | Original Paper

AAV-mediated gene delivery of the calreticulin anti-angiogenic domain inhibits ocular neovascularization

Authors: Leilei Tu, Jiang-Hui Wang, Veluchamy A. Barathi, Selwyn M. Prea, Zheng He, Jia Hui Lee, James Bender, Anna E. King, Grant J. Logan, Ian E. Alexander, Youn-Shen Bee, Ming-Hong Tai, Gregory J. Dusting, Bang V. Bui, Jingxiang Zhong, Guei-Sheung Liu

Published in: Angiogenesis | Issue 1/2018

Abstract

Ocular neovascularization is a common pathological feature in diabetic retinopathy and neovascular age-related macular degeneration that can lead to severe vision loss. We evaluated the therapeutic efficacy of a novel endogenous inhibitor of angiogenesis, the calreticulin anti-angiogenic domain (CAD180), and its functional 112-residue fragment, CAD-like peptide 112 (CAD112), delivered using a self-complementary adeno-associated virus serotype 2 (scAAV2) in rodent models of oxygen-induced retinopathy and laser-induced choroidal neovascularization. The expression of CAD180 and CAD112 was elevated in human umbilical vein endothelial cells transduced with scAAV2-CAD180 or scAAV2-CAD112, respectively, and both inhibited angiogenic activity in vitro. Intravitreal gene delivery of scAAV2-CAD180 or scAAV2-CAD112 significantly inhibited ischemia-induced retinal neovascularization in rat eyes (CAD180: 52.7% reduction; CAD112: 49.2% reduction) compared to scAAV2-mCherry, as measured in retinal flatmounts stained with isolectin B4. Moreover, the retinal structure and function were unaffected by scAAV2-CAD180 or scAAV2-CAD112, as measured by optical coherence tomography and electroretinography. Moreover, subretinal delivery of scAAV2-CAD180 or scAAV2-CAD112 significantly attenuated laser-induced choroidal neovascularization in mouse eyes compared to scAAV2-mCherry, as measured by fundus fluorescein angiography (CAD180: 62.4% reduction; CAD112: 57.5% reduction) and choroidal flatmounts (CAD180: 40.21% reduction; CAD112: 43.03% reduction). Gene delivery using scAAV2-CAD180 or scAAV2-CAD112 has significant potential as a therapeutic option for the management of ocular neovascularization.

Available only for authorised users

Campochiaro PA (2000) Retinal and choroidal neovascularization. J Cell Physiol 184(3):301–310. https://doi.org/10.1002/1097-4652(200009)184:3<301:AID-JCP3>3.0.CO;2-HCrossRefPubMed

Sapieha P, Hamel D, Shao Z, Rivera JC, Zaniolo K, Joyal JS, Chemtob S (2010) Proliferative retinopathies: angiogenesis that blinds. Int J Biochem Cell Biol 42(1):5–12. https://doi.org/10.1016/j.biocel.2009.10.006 CrossRefPubMed

Bhutto I, Lutty G (2012) Understanding age-related macular degeneration (AMD): relationships between the photoreceptor/retinal pigment epithelium/Bruch’s membrane/choriocapillaris complex. Mol Asp Med 33(4):295–317. https://doi.org/10.1016/j.mam.2012.04.005 CrossRef

Campochiaro PA (2015) Molecular pathogenesis of retinal and choroidal vascular diseases. Prog Retin Eye Res 49:67–81CrossRefPubMedPubMedCentral

Osaadon P, Fagan XJ, Lifshitz T, Levy J (2014) A review of anti-VEGF agents for proliferative diabetic retinopathy. Eye 28(5):510–520. https://doi.org/10.1038/eye.2014.13 CrossRefPubMedPubMedCentral

Writing committee for the diabetic retinopathy clinical research network, Gross JG, Glassman AR, Jampol LM, Inusah S, Aiello LP, Antoszyk AN, Baker CW, Berger BB, Bressler NM, Browning D, Elman MJ, Ferris FL, Friedman SM 3rd, Marcus DM, Melia M, Stockdale CR, Sun JK, Beck RW (2015). Panretinal photocoagulation vs intravitreous ranibizumab for proliferative diabetic retinopathy: a randomized clinical trial. JAMA 314(20):2137–2146. https://doi.org/10.1001/jama.2015.15217 CrossRef

Lally DR, Gerstenblith AT, Regillo CD (2012) Preferred therapies for neovascular age-related macular degeneration. Curr Opin Ophthalmol 23(3):182–188. https://doi.org/10.1097/ICU.0b013e328352411c CrossRefPubMed

Holz FG, Amoaku W, Donate J, Guymer RH, Kellner U, Schlingemann RO, Weichselberger A, Staurenghi G, Group SS (2011) Safety and efficacy of a flexible dosing regimen of ranibizumab in neovascular age-related macular degeneration: the SUSTAIN study. Ophthalmology 118(4):663–671. https://doi.org/10.1016/j.ophtha.2010.12.019 CrossRefPubMed

Rofagha S, Bhisitkul RB, Boyer DS, Sadda SR, Zhang K, S-US Group (2013) Seven-year outcomes in ranibizumab-treated patients in ANCHOR, MARINA, and HORIZON: a multicenter cohort study (SEVEN-UP). Ophthalmology 120(11):2292–2299. https://doi.org/10.1016/j.ophtha.2013.03.046 CrossRefPubMed

10.

Falavarjani KG, Nguyen QD (2013) Adverse events and complications associated with intravitreal injection of anti-VEGF agents: a review of literature. Eye 27(7):787–794. https://doi.org/10.1038/eye.2013.107 CrossRefPubMed

11.

Pike SE, Yao L, Jones KD, Cherney B, Appella E, Sakaguchi K, Nakhasi H, Teruya-Feldstein J, Wirth P, Gupta G, Tosato G (1998) Vasostatin, a calreticulin fragment, inhibits angiogenesis and suppresses tumor growth. J Exp Med 188(12):2349–2356CrossRefPubMedPubMedCentral

12.

Yao L, Pike SE, Setsuda J, Parekh J, Gupta G, Raffeld M, Jaffe ES, Tosato G (2000) Effective targeting of tumor vasculature by the angiogenesis inhibitors vasostatin and interleukin-12. Blood 96(5):1900–1905PubMed

13.

Sheu SJ, Bee YS, Ma YL, Liu GS, Lin HC, Yeh TL, Liou JC, Tai MH (2009) Inhibition of choroidal neovascularization by topical application of angiogenesis inhibitor vasostatin. Mol Vis 15:1897–1905PubMedPubMedCentral

14.

Bee YS, Sheu SJ, Ma YL, Lin HC, Weng WT, Kuo HM, Hsu HC, Tang CH, Liou JC, Tai MH (2010) Topical application of recombinant calreticulin peptide, vasostatin 48, alleviates laser-induced choroidal neovascularization in rats. Mol Vis 16:756–767PubMedPubMedCentral

15.

Bee YS, Tu L, Sheu SJ, Lin HC, Tang JH, Wang JH, Prea SM, Dusting GJ, Wu DC, Zhong J, Bui BV, Tai MH, Liu GS (2017) Gene delivery of calreticulin anti-angiogenic domain attenuates the development of choroidal neovascularization in rats. Hum Gene Ther. https://doi.org/10.1089/hum.2016.035 PubMed

16.

Petit L, Khanna H, Punzo C (2016) Advances in gene therapy for diseases of the eye. Hum Gene Ther 27(8):563–579CrossRefPubMedPubMedCentral

17.

Rakoczy EP, Lai CM, Magno AL, Wikstrom ME, French MA, Pierce CM, Schwartz SD, Blumenkranz MS, Chalberg TW, Degli-Esposti MA, Constable IJ (2015) Gene therapy with recombinant adeno-associated vectors for neovascular age-related macular degeneration: 1 year follow-up of a phase 1 randomised clinical trial. Lancet 386(10011):2395–2403. https://doi.org/10.1016/S0140-6736(15)00345-1 CrossRefPubMed

18.

Heier JS, Kherani S, Desai S, Dugel P, Kaushal S, Cheng SH, Delacono C, Purvis A, Richards S, Le-Halpere A, Connelly J, Wadsworth SC, Varona R, Buggage R, Scaria A, Campochiaro PA (2017) Intravitreous injection of AAV2-sFLT01 in patients with advanced neovascular age-related macular degeneration: a phase 1, open-label trial. Lancet 390(10089):50–61. https://doi.org/10.1016/s0140-6736(17)30979-0 CrossRefPubMed

19.

Pike SE, Yao L, Setsuda J, Jones KD, Cherney B, Appella E, Sakaguchi K, Nakhasi H, Atreya CD, Teruya-Feldstein J, Wirth P, Gupta G, Tosato G (1999) Calreticulin and calreticulin fragments are endothelial cell inhibitors that suppress tumor growth. Blood 94(7):2461–2468PubMed

20.

Xiao F, Wei Y, Yang L, Zhao X, Tian L, Ding Z, Yuan S, Lou Y, Liu F, Wen Y, Li J, Deng H, Kang B, Mao Y, Lei S, He Q, Su J, Lu Y, Niu T, Hou J, Huang MJ (2002) A gene therapy for cancer based on the angiogenesis inhibitor, vasostatin. Gene Ther 9(18):1207–1213. https://doi.org/10.1038/sj.gt.3301788 CrossRefPubMed

21.

Lange-Asschenfeldt B, Velasco P, Streit M, Hawighorst T, Pike SE, Tosato G, Detmar M (2001) The angiogenesis inhibitor vasostatin does not impair wound healing at tumor-inhibiting doses. J Investig Dermatol 117(5):1036–1041. https://doi.org/10.1046/j.0022-202x.2001.01519.x CrossRefPubMed

22.

Huegel R, Velasco P, De la Luz Sierra M, Christophers E, Schroder JM, Schwarz T, Tosato G, Lange-Asschenfeldt B (2007) Novel anti-inflammatory properties of the angiogenesis inhibitor vasostatin. J Investig Dermatol 127(1):65–74. https://doi.org/10.1038/sj.jid.5700484 CrossRefPubMed

23.

Tang J, Kern TS (2011) Inflammation in diabetic retinopathy. Prog Retin Eye Res 30(5):343–358CrossRefPubMedPubMedCentral

24.

Datta S, Cano M, Ebrahimi K, Wang L, Handa JT (2017) The impact of oxidative stress and inflammation on RPE degeneration in non-neovascular AMD. Prog Retin Eye Res. https://doi.org/10.1016/j.preteyeres.2017.03.002 PubMed

25.

Liu GS, Wang JH, Lee JH, Tsai PJ, Tsai HE, Sheu SJ, Lin HC, Dusting GJ, Tai MH, Bee YS (2015) Gene delivery by subconjunctival injection of adenovirus in rats: a study of local distribution transgene duration and safety. PloS one 10(12):e0143956. https://doi.org/10.1371/journal.pone.0143956 CrossRefPubMedPubMedCentral

26.

McCarty DM (2008) Self-complementary AAV vectors; advances and applications. Mol Ther J Am Soc Gene Ther 16(10):1648–1656. https://doi.org/10.1038/mt.2008.171 CrossRef

27.

Kong F, Li W, Li X, Zheng Q, Dai X, Zhou X, Boye SL, Hauswirth WW, Qu J, Pang JJ (2010) Self-complementary AAV5 vector facilitates quicker transgene expression in photoreceptor and retinal pigment epithelial cells of normal mouse. Exp Eye Res 90(5):546–554. https://doi.org/10.1016/j.exer.2010.01.011 CrossRefPubMedPubMedCentral

28.

Dalkara D, Kolstad KD, Caporale N, Visel M, Klimczak RR, Schaffer DV, Flannery JG (2009) Inner limiting membrane barriers to AAV-mediated retinal transduction from the vitreous. Mol Ther J Am Soc Gene Ther 17(12):2096–2102. https://doi.org/10.1016/j.ymthe.2016.10.008 CrossRef

29.

Takahashi K, Igarashi T, Miyake K, Kobayashi M, Yaguchi C, Iijima O, Yamazaki Y, Katakai Y, Miyake N, Kameya S, Shimada T, Takahashi H, Okada T (2017) Improved intravitreal AAV-mediated inner retinal gene transduction after surgical internal limiting membrane peeling in cynomolgus monkeys. Mol Ther J Am Soc Gene Ther 25(1):296–302CrossRef

30.

Maguire AM, Simonelli F, Pierce EA, Pugh EN Jr, Mingozzi F, Bennicelli J, Banfi S, Marshall KA, Testa F, Surace EM, Rossi S, Lyubarsky A, Arruda VR, Konkle B, Stone E, Sun J, Jacobs J, Dell’Osso L, Hertle R, Ma JX, Redmond TM, Zhu X, Hauck B, Zelenaia O, Shindler KS, Maguire MG, Wright JF, Volpe NJ, McDonnell JW, Auricchio A, High KA, Bennett J (2008) Safety and efficacy of gene transfer for Leber’s congenital amaurosis. N Engl J Med 358(21):2240–2248CrossRefPubMedPubMedCentral

31.

MacLaren RE, Groppe M, Barnard AR, Cottriall CL, Tolmachova T, Seymour L, Clark KR, During MJ, Cremers FP, Black GC, Lotery AJ, Downes SM, Webster AR, Seabra MC (2014) Retinal gene therapy in patients with choroideremia: initial findings from a phase 1/2 clinical trial. Lancet 383(9923):1129–1137CrossRefPubMedPubMedCentral

32.

Lai CM, Estcourt MJ, Himbeck RP, Lee SY, Yew-San Yeo I, Luu C, Loh BK, Lee MW, Barathi A, Villano J, Ang CL, van der Most RG, Constable IJ, Dismuke D, Samulski RJ, Degli-Esposti MA, Rakoczy EP (2012) Preclinical safety evaluation of subretinal AAV2.sFlt-1 in non-human primates. Gene Ther 19(10):999–1009. https://doi.org/10.1038/gt.2011.169 CrossRefPubMed

33.

Dalkara D, Byrne LC, Klimczak RR, Visel M, Yin L, Merigan WH, Flannery JG, Schaffer DV (2013) In vivo-directed evolution of a new adeno-associated virus for therapeutic outer retinal gene delivery from the vitreous. Sci Transl Med 5(189):189ra176. https://doi.org/10.1126/scitranslmed.3005708 CrossRef

34.

Ip MS, Scott IU, Brown GC, Brown MM, Ho AC, Huang SS, Recchia FM (2008) Anti-vascular endothelial growth factor pharmacotherapy for age-related macular degeneration: a report by the American Academy of Ophthalmology. Ophthalmology 115(10):1837–1846. https://doi.org/10.1016/j.ophtha.2008.08.012 CrossRefPubMed

35.

Nguyen QD, Brown DM, Marcus DM, Boyer DS, Patel S, Feiner L, Gibson A, Sy J, Rundle AC, Hopkins JJ, Rubio RG, Ehrlich JS (2012) Ranibizumab for diabetic macular edema: results from 2 phase III randomized trials: RISE and RIDE. Ophthalmology 119(4):789–801. https://doi.org/10.1016/j.ophtha.2011.12.039 CrossRefPubMed

36.

Curtis LH, Hammill BG, Schulman KA, Cousins SW (2010) Risks of mortality, myocardial infarction, bleeding, and stroke associated with therapies for age-related macular degeneration. Arch Ophthalmol 128(10):1273–1279. https://doi.org/10.1001/archophthalmol.2010.223 (Chicago, Ill: 1960) CrossRefPubMed

37.

Avery RL, Gordon GM (2016) Systemic safety of prolonged monthly anti-vascular endothelial growth factor therapy for diabetic macular edema: a systematic review and meta-analysis. JAMA Ophthalmol 134(1):21–29. https://doi.org/10.1001/jamaophthalmol.2015.4070 CrossRefPubMed

38.

McCarty DM, Monahan PE, Samulski RJ (2001) Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently of DNA synthesis. Gene Ther 8(16):1248–1254. https://doi.org/10.1038/sj.gt.3301514 CrossRefPubMed

39.

Liu GS, Tsai HE, Weng WT, Liu LF, Weng CH, Chuang MR, Lam HC, Wu CS, Tee R, Wen ZH, Howng SL, Tai MH (2011) Systemic pro-opiomelanocortin expression induces melanogenic differentiation and inhibits tumor angiogenesis in established mouse melanoma. Hum Gene Ther 22(3):325–335. https://doi.org/10.1089/hum.2010.090 CrossRefPubMed

40.

Carpentier G, Martinelli M, Courty J, Cascone I (2012) Angiogenesis analyzer for ImageJ. In: 4th ImageJ User and developer conference proceedings, Mondorf-les-Bains, Luxembourg

41.

Deliyanti D, Miller AG, Tan G, Binger KJ, Samson AL, Wilkinson-Berka JL (2012) Neovascularization is attenuated with aldosterone synthase inhibition in rats with retinopathy. Hypertension 59(3):607–613. https://doi.org/10.1161/hypertensionaha.111.188136 CrossRefPubMed

42.

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25(4):402–408. https://doi.org/10.1006/meth.2001.1262 CrossRefPubMed

43.

Connor KM, Krah NM, Dennison RJ, Aderman CM, Chen J, Guerin KI, Sapieha P, Stahl A, Willett KL, Smith LE (2009) Quantification of oxygen-induced retinopathy in the mouse: a model of vessel loss, vessel regrowth and pathological angiogenesis. Nat Protoc 4(11):1565–1573CrossRefPubMedPubMedCentral

44.

Nivison-Smith L, Zhu Y, Whatham A, Bui BV, Fletcher EL, Acosta ML, Kalloniatis M (2014) Sildenafil alters retinal function in mouse carriers of retinitis pigmentosa. Exp Eye Res 128:43–56. https://doi.org/10.1016/j.exer.2014.08.014 CrossRefPubMed

Title: AAV-mediated gene delivery of the calreticulin anti-angiogenic domain inhibits ocular neovascularization
Authors: Leilei Tu
Jiang-Hui Wang
Veluchamy A. Barathi
Selwyn M. Prea
Zheng He
Jia Hui Lee
James Bender
Anna E. King
Grant J. Logan
Ian E. Alexander
Youn-Shen Bee
Ming-Hong Tai
Gregory J. Dusting
Bang V. Bui
Jingxiang Zhong
Guei-Sheung Liu
Publication date: 01-02-2018
Publisher: Springer Netherlands
Published in: Angiogenesis / Issue 1/2018
Print ISSN: 0969-6970
Electronic ISSN: 1573-7209
DOI: https://doi.org/10.1007/s10456-017-9591-4

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Heart Failure Video

Year in Review: Heart failure and cardiomyopathies

Watch now

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits

STEP AAG HeroTeaserCollection image/© Tarek / Generated with AI / Stock.adobe.com, ACC 2024 Pediatric and Congenital Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Pulmonary Vascular Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Valvular Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 HF and Cardiomyopathies: Year in Review/© 2024 American College of Cardiology, Woman out walking/© Seventyfour / stock.adobe.com (symbolic image with model), Woman checking smartwatch /© saltdium / stock.adobe.com (symbolic image with model), Cardiac catheterization/© Damian / stock.adobe.com

At a glance: The STEP trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2018

Antiangiogenesis and medical therapy failure in intracranial atherosclerosis

12th International HHT Scientific Conference

Noninvasive induction of angiogenesis in tissues by external suction: sequential optimization for use in reconstructive surgery

Mechanisms of angiogenesis in microbe-regulated inflammatory and neoplastic conditions

Notch signaling controls sprouting angiogenesis of endometriotic lesions

Dimethylarginine dimethylaminohydrolase-1 (DDAH1) is frequently upregulated in prostate cancer, and its overexpression conveys tumor growth and angiogenesis by metabolizing asymmetric dimethylarginine (ADMA)

Highlights from the ACC 2024 Congress

ACC 2024 Congress Coverage