目的:使用条件重编程(CR)建立和表征头颈部鳞状细胞癌(HNSCC)培养物的多样化文库。
方法:患者参加了IRB批准的方案,以使用CR方法产生肿瘤细胞培养物。收集肿瘤和血液样本并记录临床信息。成功的CR培养物针对具有短串联重复基因分型的库存参考肿瘤进行了验证。细胞形态学用照片记录存档。评估临床和人口统计学因素与成功建立CR文化的关联。人乳头瘤病毒(HPV)基因分型,克隆生存,MTT测定,球体生长,在选定的培养物中进行全外显子组测序。
结果:纳入了44例患者,31(70%)成功的CR文化,32%来自确定为黑人的患者,61%来自西班牙裔。所有主要的头颈部疾病部位都有代表,包括15例(48%)口腔癌和8例(26%)p16阳性口咽癌。西班牙裔种族和第一原发性肿瘤(vs.第二原发或复发性肿瘤)与成功的CR培养显着相关。HPV表达在CR培养物中是保守的,包括CR-024,其携带新的HPV-69血清型。CR培养物用于使用MTT测定法测试顺铂反应。先前的工作还表明,这些模型可用于评估对辐射的反应,并可移植到小鼠模型中。全外显子组测序表明CR培养物保留了肿瘤突变负荷和驱动突变。
结论:CR培养在增殖HNSCC细胞方面非常成功。这项研究包括来自代表性不足的少数群体的高比例患者。
方法:不适用喉镜,2024.
OBJECTIVE: To establish and characterize a diverse library of head and neck squamous cell cancer (HNSCC) cultures using conditional reprogramming (CR).
METHODS: Patients enrolled on an IRB-approved protocol to generate tumor cell cultures using CR methods. Tumor and blood samples were collected and clinical information was recorded. Successful CR cultures were validated against banked reference tumors with short tandem repeat genotyping. Cell morphology was archived with photodocumentation. Clinical and demographic factors were evaluated for associations with successful establishment of CR culture. Human papilloma virus (HPV) genotyping, clonogenic survival, MTT assays, spheroid growth, and whole exome sequencing were carried out in selected cultures.
RESULTS: Forty four patients were enrolled, with 31 (70%) successful CR cultures, 32% derived from patients who identified as Black and 61% as Hispanic. All major head and neck disease sites were represented, including 15 (48%) oral cavity and 8 (26%) p16-positive oropharynx cancers. Hispanic ethnicity and first primary tumors (vs. second primary or recurrent tumors) were significantly associated with successful CR culture. HPV expression was conserved in CR cultures, including CR-024, which carried a novel HPV-69 serotype. CR cultures were used to test cisplatin responses using MTT assays. Previous work has also demonstrated these models can be used to assess response to radiation and can be engrafted in mouse models. Whole exome sequencing demonstrated that CR cultures preserved tumor mutation burden and driver mutations.
CONCLUSIONS: CR culture is highly successful in propagating HNSCC cells. This study included a high proportion of patients from underrepresented minority groups.
METHODS: Not Applicable Laryngoscope, 134:2748-2756, 2024.