Functional Characterization of NRAS-G12D in a Microfluidic 3D ECM Platform
Eunu Baek, Elliot Hoyoung Nam, Eunice Jiwoo Kang, Soo Young Pee, Aiden Seunghwan Chee, Eileen Byon, Woo Joo Kang, Younho Aiden Park, Andrew Suh, Si Hyun Lee, Seunghu Lee, Minjun Hwang
iGEM KOREA-CX, Seoul, South Korea
Publication date: July 10, 2026
iGEM KOREA-CX, Seoul, South Korea
Publication date: July 10, 2026
DOI: http://doi.org/10.34614/JIYRC2026I06
ABSTRACT
Multiple myeloma progression is shaped not only by oncogenic mutations in the RAS pathway but also by physical and biochemical features of the bone marrow niche. Conventional 2D cultures have limited capacity to capture genotype-microenvironment interactions, and animal models have translational limitations. Here, we engineered NIH3T3 cells expressing control, NRAS-WT, or NRAS-G12D constructs and integrated them into a PDMS-based lab-on-a-chip platform containing a Matrigel-derived 3D matrix and an 8 μm porous membrane. NRAS-G12D expression enhanced proliferation, accelerated wound closure, and increased infiltration under 3D conditions. Under U0126 treatment, NRAS-G12D cells maintained higher viability than control and NRAS-WT cells, indicating a genotype-dependent drug-response phenotype under the tested conditions. These results show that the platform enables controlled comparison of mutation-associated behaviors within a bone-marrow-inspired 3D microenvironment and should be interpreted as a proof-of-concept system for mechanistic studies rather than a validated predictor of clinical therapeutic response.
Multiple myeloma progression is shaped not only by oncogenic mutations in the RAS pathway but also by physical and biochemical features of the bone marrow niche. Conventional 2D cultures have limited capacity to capture genotype-microenvironment interactions, and animal models have translational limitations. Here, we engineered NIH3T3 cells expressing control, NRAS-WT, or NRAS-G12D constructs and integrated them into a PDMS-based lab-on-a-chip platform containing a Matrigel-derived 3D matrix and an 8 μm porous membrane. NRAS-G12D expression enhanced proliferation, accelerated wound closure, and increased infiltration under 3D conditions. Under U0126 treatment, NRAS-G12D cells maintained higher viability than control and NRAS-WT cells, indicating a genotype-dependent drug-response phenotype under the tested conditions. These results show that the platform enables controlled comparison of mutation-associated behaviors within a bone-marrow-inspired 3D microenvironment and should be interpreted as a proof-of-concept system for mechanistic studies rather than a validated predictor of clinical therapeutic response.
