Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing
Abstract Background Sickle cell disease (SCD) is a disorder of red blood cells (RBCs) expressing abnormal hemoglobin-S (HbS) due to genetic inheritance of homologous HbS gene. However, people with the sickle cell trait (SCT) carry a single allele of HbS and do not usually suffer from SCD symptoms, t...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2017-06-01
|
| Series: | Journal of Hematology & Oncology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13045-017-0489-9 |
| _version_ | 1819135852125618176 |
|---|---|
| author | Jianguo Wen Wenjing Tao Suyang Hao Youli Zu |
| author_facet | Jianguo Wen Wenjing Tao Suyang Hao Youli Zu |
| author_sort | Jianguo Wen |
| collection | DOAJ |
| description | Abstract Background Sickle cell disease (SCD) is a disorder of red blood cells (RBCs) expressing abnormal hemoglobin-S (HbS) due to genetic inheritance of homologous HbS gene. However, people with the sickle cell trait (SCT) carry a single allele of HbS and do not usually suffer from SCD symptoms, thus providing a rationale to treat SCD. Methods To validate gene therapy potential, hematopoietic stem cells were isolated from the SCD patient blood and treated with CRISPR/Cas9 approach. To precisely dissect genome-editing effects, erythroid progenitor cells were cloned from single colonies of CRISPR-treated cells and then expanded for simultaneous gene, protein, and cellular function studies. Results Genotyping and sequencing analysis revealed that the genome-edited erythroid progenitor colonies were converted to SCT genotype from SCD genotype. HPLC protein assays confirmed reinstallation of normal hemoglobin at a similar level with HbS in the cloned genome-edited erythroid progenitor cells. For cell function evaluation, in vitro RBC differentiation of the cloned erythroid progenitor cells was induced. As expected, cell sickling assays indicated function reinstitution of the genome-edited offspring SCD RBCs, which became more resistant to sickling under hypoxia condition. Conclusions This study is an exploration of genome editing of SCD HSPCs. |
| first_indexed | 2024-12-22T10:25:40Z |
| format | Article |
| id | doaj.art-5edf25f97b09426b89722d533f478699 |
| institution | Directory Open Access Journal |
| issn | 1756-8722 |
| language | English |
| last_indexed | 2024-12-22T10:25:40Z |
| publishDate | 2017-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Hematology & Oncology |
| spelling | doaj.art-5edf25f97b09426b89722d533f4786992022-12-21T18:29:30ZengBMCJournal of Hematology & Oncology1756-87222017-06-0110111110.1186/s13045-017-0489-9Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editingJianguo Wen0Wenjing Tao1Suyang Hao2Youli Zu3Department of Pathology and Genomic Medicine, Houston Methodist Hospital, Houston Methodist Research InstituteDepartment of Leukemia, The University of Texas M.D. Anderson Cancer CenterDepartment of Pathology and Genomic Medicine, Houston Methodist Hospital, Houston Methodist Research InstituteDepartment of Pathology and Genomic Medicine, Houston Methodist Hospital, Houston Methodist Research InstituteAbstract Background Sickle cell disease (SCD) is a disorder of red blood cells (RBCs) expressing abnormal hemoglobin-S (HbS) due to genetic inheritance of homologous HbS gene. However, people with the sickle cell trait (SCT) carry a single allele of HbS and do not usually suffer from SCD symptoms, thus providing a rationale to treat SCD. Methods To validate gene therapy potential, hematopoietic stem cells were isolated from the SCD patient blood and treated with CRISPR/Cas9 approach. To precisely dissect genome-editing effects, erythroid progenitor cells were cloned from single colonies of CRISPR-treated cells and then expanded for simultaneous gene, protein, and cellular function studies. Results Genotyping and sequencing analysis revealed that the genome-edited erythroid progenitor colonies were converted to SCT genotype from SCD genotype. HPLC protein assays confirmed reinstallation of normal hemoglobin at a similar level with HbS in the cloned genome-edited erythroid progenitor cells. For cell function evaluation, in vitro RBC differentiation of the cloned erythroid progenitor cells was induced. As expected, cell sickling assays indicated function reinstitution of the genome-edited offspring SCD RBCs, which became more resistant to sickling under hypoxia condition. Conclusions This study is an exploration of genome editing of SCD HSPCs.http://link.springer.com/article/10.1186/s13045-017-0489-9CRISPR/Cas9 genome editingHematopoietic stem/progenitor cell coloniesSickle cell disease |
| spellingShingle | Jianguo Wen Wenjing Tao Suyang Hao Youli Zu Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing Journal of Hematology & Oncology CRISPR/Cas9 genome editing Hematopoietic stem/progenitor cell colonies Sickle cell disease |
| title | Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing |
| title_full | Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing |
| title_fullStr | Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing |
| title_full_unstemmed | Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing |
| title_short | Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing |
| title_sort | cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post crispr genome editing |
| topic | CRISPR/Cas9 genome editing Hematopoietic stem/progenitor cell colonies Sickle cell disease |
| url | http://link.springer.com/article/10.1186/s13045-017-0489-9 |
| work_keys_str_mv | AT jianguowen cellularfunctionreinstitutionofoffspringredbloodcellsclonedfromthesicklecelldiseasepatientbloodpostcrisprgenomeediting AT wenjingtao cellularfunctionreinstitutionofoffspringredbloodcellsclonedfromthesicklecelldiseasepatientbloodpostcrisprgenomeediting AT suyanghao cellularfunctionreinstitutionofoffspringredbloodcellsclonedfromthesicklecelldiseasepatientbloodpostcrisprgenomeediting AT youlizu cellularfunctionreinstitutionofoffspringredbloodcellsclonedfromthesicklecelldiseasepatientbloodpostcrisprgenomeediting |