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Original research
Quantification of human embryonic ζ-globin chains in Southeast Asian deletion (--SEA) carriers
  1. Yuhua Ye1,2,3,
  2. Guoying Sun4,
  3. Zhe Ren4,
  4. Yidan Liang1,2,3,
  5. Hualei Luo1,2,3,
  6. Peng Lin1,2,3,
  7. Xingmin Wang1,2,3,
  8. Zejun Dong1,2,3,
  9. Li Huang1,2,3,
  10. Lang Qin1,2,3,
  11. Wenfang Yu5,
  12. Ge Wang6,
  13. Yuqiu Zhou6,
  14. Jia Tang7,
  15. Jiwu Lou8,
  16. Yanhui Liu8,
  17. Xianqi Zeng9,
  18. Yajun Chen9,
  19. Yihong Li10,
  20. Qianqian Zhang1,2,3,
  21. Jin Huang1,2,3,
  22. Ping Zhu11,
  23. Liang Lin4,
  24. Xinhua Zhang12,
  25. Xiangmin Xu1,2,3
  1. 1 Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, People's Republic of China
  2. 2 Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, People's Republic of China
  3. 3 Guangdong Genetics Testing Engineering Research Center, Guangzhou, People's Republic of China
  4. 4 BGI Genomics, BGI-Shenzhen, Shenzhen, Guangdong, People's Republic of China
  5. 5 Department of Blood Transfusion, Shanghai General Hospital, Shanghai, People's Republic of China
  6. 6 Department of Clinical Laboratory, Zhuhai Municipal Maternal and Child Healthcare Hospital, Zhuhai, Guangdong, People's Republic of China
  7. 7 NHC Key Laboratory of Male Reproduction and Genetics, Guangdong Provincial Reproductive Science Institute, Guangzhou, People's Republic of China
  8. 8 Dongguan Institute of Reproduction and Genetics, Dongguan Maternal and Children Health Hospital, Dongguan, People's Republic of China
  9. 9 Women and Children's Health Hospital of Shaoguan, Shaoguan, Guangdong, People's Republic of China
  10. 10 Department of Gynecology and Obstetrics, Southern Medical University, Guangzhou, People's Republic of China
  11. 11 Department of Immunology, Southern Medical University, Guangzhou, People's Republic of China
  12. 12 Department of Hematology, 923rd Hospital of the People's Liberation Army, Nanning, Guangxi, People's Republic of China
  1. Correspondence to Dr Xiangmin Xu, Department of Medical Genetics, Southern Medical University, Guangzhou, China; xixm{at}smu.edu.cn

Abstract

Aims Reactivation of embryonic ζ-globin is a promising strategy for genetic treatment of α-thalassaemia. However, quantification of ζ-globin as a quantitative trait in α-thalassaemia carriers and patients remains incompletely understood. In this study, we aimed to set up a reliable approach for the quantification of ζ-globin in α-thalassaemia carriers, followed by a population study to investigate its expression patterns.

Methods ζ-globin was purified as monomers from cord blood haemolysate of a Hb Bart’s fetus, followed by absolute protein quantification, which was then tested by in-house ELISA system and introduced as protein standard. It was then used for large-scale quantification in peripheral blood samples from 6179 individuals. Finally, liquid chromatography-tandem mass spectrometry (LC-MS/MS) introduced as an independent validating approach by measuring ζ-globin expression in a second cohort of 141-SEA/αα carriers.

Results The ELISA system was proved sensitive in distinguishing individuals with varied extent of ζ-globin. Large scale quantitative study of this --SEA/αα carrier cohort indicated the high diversity of ζ-globin expression ranging from 0.00155 g/L to 1.48778 g/L. Significant positive correlation between ELISA and LC-MS/MS (R=0.400, p<0.001) was observed and it is more sensitive in distinguishing the samples with extreme expression of ζ-globin (R=0.650, p<0.001).

Conclusion Our study has reported reliable approaches for the quantification of ζ-globin and presented the expression patterns of ζ-globin among the --SEA/αα carrier population, which might lay a foundation on subsequent genotype–phenotype studies on mechanisms of delayed haemoglobin switch in α-thalassaemia.

  • thalassemia
  • hemoglobinopathies
  • embryonic and fetal development

Data availability statement

Data are available on reasonable request.

https://doi.org/10.1136/jcp-2022-208159

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    Data availability statement

    Data are available on reasonable request.

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    Footnotes

    • Handling editor Tahir S Pillay.

    • YY and GS contributed equally.

    • Contributors YY, LL and XX designed the study. YY, GS, ZR and YaL performed experiments and analyzed the data. HL, PL, XW, ZD, QZ, PZ and JH performed experiments. XZ, YC, JT, JL, YaL, XZ and YiL collected and analysed clinical data. YY, GS, ZR, LL and XX wrote the manuscript. XX is responsible for the overall content as guarantor. All authors read and approved the final manuscript.

    • Funding This study was supported by National Natural Science Foundation of China (Grant No. 31871265) and Shanghai Municipal Science and Technology Major Project (Grant No. 2017SHZDZX01).

    • Competing interests None declared.

    • Provenance and peer review Not commissioned; externally peer reviewed.

    • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.