Establishment of a patient-derived organoid model and living biobank for nasopharyngeal carcinoma

Abstract

Background Recurrent nasopharyngeal carcinoma (NPC) remains a major challenge for clinicians and scientists. Tumor organoid is a revelational disease model that highly resembled the heterogeneity and histopathological characteristics of original tumors. This study aimed to optimize the modeling process of patient-derived NPC organoids (NPCOs), and establish a living-biobank of NPCs to study the mechanism and explore the more effective treatment of the disease. Methods Sixty-two fresh NPC tissue samples and 15 normal mucosa samples were collected for 3-dimensional (3D) organoid culture. The organoids were confirmed using hematoxylin and eosin assays. The expression levels of CD133, CD44, BMI-1 , and Epstein-Barr virus (EBV)-encoded small RNAs (EBERs) were detected by immunohistochemistry (IHC) and in situ hybridization (ISH). Recurrent NPCOs were frozen in liquid nitrogen for 6 months and then resuscitated and passaged. Results We identified a novel two-step enzymatic strategy for the treatment of NPC and nasal mucosa specimens and an optimal medium for culturing NPCOs and nasal mucosa organoids (NMOs). Organoid cultures were generated from 34 primary NPC samples, 28 recurrent NPC samples, and 15 normal mucosa samples. The success rates for primary NPCO, recurrent NPCO, and NMO formation were 47.06%, 81.25%, and 86.5%, respectively. All the NPCOs were EBER positive and CK7 negative. Recurrent NPCOs had higher expressions of stem cell markers, including BMI-1, CD44 , and CD133 . Additionally, recurrent NPCOs could be cultured to passage 4 and frozen and revived repeatedly, while primary NPCOs were challenging to culture. Conclusions In summary, we successfully established a living biobank using the NPCO model, which has enormous potential in basic and clinical research on NPC.

Targeted therapy for head and neck cancer: signaling pathways and clinical studies

“…228 As mentioned previously, the PDX model provides a novel approach for identifying suitable alterations in tumor tissue to find promising treatment methods. [229][230][231][232] Recently, this model has been considered an ideal preclinical model for investigating targeted drugs. These PDX models maintained the genetic background in immunodeficient mice.…”

Tumor organoid biobank-new platform for medical research

“…The high recurrence rate is responsible for the poor prognosis for most nasopharyngeal cancers 53 . Wang et al 54 established a PDTO biobank of 39 primary and recurrent nasopharyngeal carcinomas and found that all nasopharyngeal carcinomas carried Epstein-Barr virus and kept the virus expanding. Furthermore, their stem cell markers were expressed more in recurrent organoids than in primary.…”

Tumor organoid model of colorectal cancer (Review)

“…To date, researchers have established live organoids for various tumors, including locally advanced rectal cancer ( 82 ), CRC, liver metastasis of CRC ( 80 ), breast cancer ( 83 ), bladder cancer ( 84 ), nasopharyngeal cancer ( 85 ), pancreatic cancer ( 86 ), non-small cell lung cancer ( 87 ), gastric cancer ( 88 ) and glioblastoma ( 89 ). For the first time, van de Wetering et al ( 41 ) successfully established a living biobank for CRC, including 22 tumor organoids and 19 normal adjacent organoids from 20 patients.…”