Non-Invasive Prenatal Diagnosis of Chromosomal and Monogenic Disease by a Novel Bioinspired Micro–Nanochip for Isolating Fetal Nucleated Red Blood Cells

Naiqi Li,1,2,* Yue Sun,3,4,* Lin Cheng,1,5,6 Chun Feng,7 Yifan Sun,1,5,6 Saisai Yang,1,5 Yuqi Shao,1,6 Xing-Zhong Zhao,3 Yuanzhen Zhang1,5,6 1Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, 430071, People’s Republic of China; 2Genetics and Prenatal Diagnosis Center, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, 450000, People’s Republic of China; 3Key Laboratory of Artificial Micro- and Nano-Structures of Ministry of Education, School of Physics and Technology, Wuhan University, Wuhan, 430072, People’s Republic of China; 4School of Physics and Electronic Engineering, Xinyang Normal University, Xinyang, 464000, People’s Republic of China; 5Hubei Clinical Research Center for Prenatal Diagnosis and Birth Health, Wuhan, 430071, People’s Republic of China; 6Wuhan Clinical Research Center for Reproductive Science and Birth Health, Wuhan, 430071, People’s Republic of China; 7Department of Gynecology, Maternal and Child Health Hospital of Hubei Province, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430072, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xing-Zhong Zhao, Key Laboratory of Artificial Micro- and Nano-Structures of Ministry of Education, School of Physics and Technology, Wuhan University, Wuhan, People’s Republic of China, Email [email protected] Yuanzhen Zhang, Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University, Wuhan, People’s Republic of China, Email [email protected]: Fetal nucleated red blood cells (fNRBCs) in the peripheral blood of pregnant women contain comprehensive fetal genetic information, making them an ideal target for non-invasive prenatal diagnosis (NIPD). However, challenges in identifying, enriching, and detecting fNRBCs limit their diagnostic potential.Methods: To overcome these obstacles, we developed a novel biomimetic chip, replicating the micro-nano structure of red rose petals on polydimethylsiloxane (PDMS). The surface was modified with gelatin nanoparticles (GNPs) and affinity antibodies to enhance cell adhesion and facilitate specific cell identification. We subsequently investigated the chip’s characteristics, along with its in vitro capture and release system, and conducted further experiments using peripheral blood samples from pregnant women.Results: In the cell line capture and release assay, the chip achieved a cell capture efficiency of 90.4%. Following metalloproteinase-9 (MMP-9) enzymatic degradation, the release efficiency was 84.08%, with cell viability at 85.97%. Notably, fNRBCs can be captured from the peripheral blood of pregnant women as early as 7 weeks of gestation. We used these fNRBCs to diagnose a case of single-gene disease and instances of chromosomal aneuploidies, yielding results consistent with those obtained from amniotic fluid punctures.Conclusion: This novel chip not only enables efficient enrichment of fNRBCs for NIPD but also extends the diagnostic window for genetic and developmental disorders to as early as 7 weeks of gestation, potentially allowing for earlier interventions. By improving the accuracy and reliability of NIPD, this technology could reduce reliance on invasive diagnostic techniques, offering a new pathway for diagnosing fetal genetic conditions in clinical practice.Keywords: non-invasive prenatal diagnosis, fetal nucleated red blood cells, nanostructure microchip, chromosomal aneuploidy, monogenic disease