Structural basis of receptor usage by the engineered capsid AAV-PHP.eB
Adeno-associated virus serotype 9 (AAV9) is a promising gene therapy vector for treating neurodegenerative diseases due to its ability to penetrate the blood-brain barrier. PHP.eB was engineered from AAV9 by insertion of a 7-amino acid peptide and point mutation of neighboring residues, thereby enha...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050122001036 |
| _version_ | 1811216795581808640 |
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| author | Seongmin Jang Hao K. Shen Xiaozhe Ding Timothy F. Miles Viviana Gradinaru |
| author_facet | Seongmin Jang Hao K. Shen Xiaozhe Ding Timothy F. Miles Viviana Gradinaru |
| author_sort | Seongmin Jang |
| collection | DOAJ |
| description | Adeno-associated virus serotype 9 (AAV9) is a promising gene therapy vector for treating neurodegenerative diseases due to its ability to penetrate the blood-brain barrier. PHP.eB was engineered from AAV9 by insertion of a 7-amino acid peptide and point mutation of neighboring residues, thereby enhancing potency in the central nervous system. Here, we report a 2.24-Å resolution cryo-electron microscopy structure of PHP.eB, revealing conformational differences from other 7-mer insertion capsid variants. In PHP.eB, the 7-mer loop adopts a bent conformation, mediated by an interaction between engineered lysine and aspartate residues. Further, we identify PKD2 as the main AAV receptor (AAVR) domain recognizing both AAV9 and PHP.eB and find that the PHP.eB 7-mer partially destabilizes this interaction. Analysis of previously reported AAV structures together with our pull-down data demonstrate that the 7-mer topology determined by the lysine-aspartate interaction dictates AAVR binding strength. Our results suggest that PHP.eB’s altered tropism may arise from both an additional interaction with LY6A and weakening of its AAVR interaction. Changing the insertion length, but not sequence, modifies PKD2 binding affinity, suggesting that a steric clash impedes AAVR binding. This research suggests improved library designs for future AAV selections to identify non-LY6A-dependent vectors and modulate AAVR interaction strength. |
| first_indexed | 2024-04-12T06:45:17Z |
| format | Article |
| id | doaj.art-df4fd9859c1b4dfeae3dfa8b780af72e |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-04-12T06:45:17Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-df4fd9859c1b4dfeae3dfa8b780af72e2022-12-22T03:43:37ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012022-09-0126343354Structural basis of receptor usage by the engineered capsid AAV-PHP.eBSeongmin Jang0Hao K. Shen1Xiaozhe Ding2Timothy F. Miles3Viviana Gradinaru4Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USADivision of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USADivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USADivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USADivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA; Corresponding author Viviana Gradinaru, PhD, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.Adeno-associated virus serotype 9 (AAV9) is a promising gene therapy vector for treating neurodegenerative diseases due to its ability to penetrate the blood-brain barrier. PHP.eB was engineered from AAV9 by insertion of a 7-amino acid peptide and point mutation of neighboring residues, thereby enhancing potency in the central nervous system. Here, we report a 2.24-Å resolution cryo-electron microscopy structure of PHP.eB, revealing conformational differences from other 7-mer insertion capsid variants. In PHP.eB, the 7-mer loop adopts a bent conformation, mediated by an interaction between engineered lysine and aspartate residues. Further, we identify PKD2 as the main AAV receptor (AAVR) domain recognizing both AAV9 and PHP.eB and find that the PHP.eB 7-mer partially destabilizes this interaction. Analysis of previously reported AAV structures together with our pull-down data demonstrate that the 7-mer topology determined by the lysine-aspartate interaction dictates AAVR binding strength. Our results suggest that PHP.eB’s altered tropism may arise from both an additional interaction with LY6A and weakening of its AAVR interaction. Changing the insertion length, but not sequence, modifies PKD2 binding affinity, suggesting that a steric clash impedes AAVR binding. This research suggests improved library designs for future AAV selections to identify non-LY6A-dependent vectors and modulate AAVR interaction strength.http://www.sciencedirect.com/science/article/pii/S2329050122001036adeno-associated virusAAVAAVRLY6APHP.eBblood-brain barrier |
| spellingShingle | Seongmin Jang Hao K. Shen Xiaozhe Ding Timothy F. Miles Viviana Gradinaru Structural basis of receptor usage by the engineered capsid AAV-PHP.eB Molecular Therapy: Methods & Clinical Development adeno-associated virus AAV AAVR LY6A PHP.eB blood-brain barrier |
| title | Structural basis of receptor usage by the engineered capsid AAV-PHP.eB |
| title_full | Structural basis of receptor usage by the engineered capsid AAV-PHP.eB |
| title_fullStr | Structural basis of receptor usage by the engineered capsid AAV-PHP.eB |
| title_full_unstemmed | Structural basis of receptor usage by the engineered capsid AAV-PHP.eB |
| title_short | Structural basis of receptor usage by the engineered capsid AAV-PHP.eB |
| title_sort | structural basis of receptor usage by the engineered capsid aav php eb |
| topic | adeno-associated virus AAV AAVR LY6A PHP.eB blood-brain barrier |
| url | http://www.sciencedirect.com/science/article/pii/S2329050122001036 |
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