Biology subjects

Hawkins, S. M.

Publications and source records attributed to Hawkins, S. M..

3 recordsLinked to original sources

bioRxiv2025

Establishment and Characterization of a New Immortalized Human Adenomyosis Epithelial-Like Cell Line, tAEC21

Adenomyosis occurs when endometrial glands and stroma grow within the uterine myometrium. Adenomyosis, as a clinically impactful disease, causes significant pelvic pain and heavy menstrual bleeding. Adenomyosis remains understudied due to the paucity of translational research tools and model systems. This study aimed to create a telomerase-transformed epithelial-like cell line derived from the eutopic endometrium of a subject with focal adenomyosis. De-identified endometrium was processed via mechanical and enzymatic digestion. Epithelial and stromal populations were separated via selective adhesion, followed by fluorescence-activated cell sorting with an epithelial cellular adhesion molecule (EpCAM). EpCAM+ cells were efficiently immortalized with the human telomerase reverse transcriptase gene. Analyses confirmed that the cells were human, without mycoplasma contamination, and exhibited a unique 16-marker short tandem repeat profile. Cytogenetic analysis on G-banded metaphase spreads indicated polyploidy with multiple chromosomal rearrangements. The line, designated as tAEC21, expressed epithelial markers cytokeratin-5 and N-cadherin but not stromal marker CD10. Cells avidly responded to tumor necrosis factor-alpha stimulation by upregulating interleukin-6, C-X-C motif chemokine ligand 8, C-C motif chemokine ligand 2, and mucin 1 gene expression. In a heterotypic, three-dimensional spheroid model, tAEC21 displayed a biologically relevant pattern by assembling into an epithelial shell around the stromal cell core. In two-dimensional monolayers, tAEC21 cells were negative for estrogen and progesterone receptors, while as 3D spheroids, tAEC21 exhibited strong positivity for estrogen but not progesterone receptors. This new epithelial-like, adenomyosis-derived cell line, tAEC21, will be an impactful research resource.

cell biology↗

bioRxiv2020

Vaginal squamous cell carcinoma develops in mice with Arid1a loss and gain of oncogenic Kras

Recent sequencing studies showed that loss-of-function mutations in ARID1A (AT-rich interactive domain 1a) were enriched in gynecologic malignancies. However, multiple mouse models with deletion of Arid1a did not exhibit gynecologic malignancy. Oncogenic KRAS mutations are a common finding in endometrial cancers. However, expression of oncogenic Kras (KrasG12D) in the uterus was not sufficient to develop endometrial cancer. These results suggest that both ARID1A deletion and oncogenic KRAS require additional hits before driving gynecologic malignancy. To determine the role of the combination effects of deletion of Arid1a and oncogenic Kras, Arid1aflox/flox mice were crossed to KrasLox-Stop-Lox-G12D/+ mice using progesterone receptor Cre (PgrCre/+). Survival studies, histology, and immunohistochemistry were used to characterize the phenotype. Hormone dependence was evaluated by ovarian hormone depletion and estradiol replacement. Arid1aflox/flox; KrasLox-Stop-Lox-G12D/+; PgrCre/+ (AKP) mice exhibited early euthanasia due to large vaginal tumors, which were invasive squamous cell carcinoma. Younger mice exhibited precancerous intraepithelial lesions that progressed to invasive squamous cell carcinoma with age. Immunohistochemistry supported the pathological diagnosis with abnormal expression and localization of cytokeratin 5, tumor protein P63, cyclin dependent kinase inhibitor 2A (CDKN2A or p16), and marker of proliferation Ki-67. Vaginal lesions in AKP mice were hormone dependent. Ovarian hormone deletion in AKP mice resulted in atrophic vaginal epithelium without evidence of vaginal tumors. Estradiol replacement in ovarian hormone depleted AKP mice resulted in lesions that resembled the squamous cell carcinoma in intact mice. AKP mice did not develop endometrial cancer. Arid1a deletion with KrasG12D expression drives invasive vaginal squamous cell carcinoma. This mouse can be used to study the transition from benign precursor lesions into invasive vaginal squamous cell carcinoma offering insights into progression.

cancer biology↗

bioRxiv2020

Building a virtual summer research experience in cancer for high school and early undergraduate students: lessons from the COVID-19 pandemic

BackgroundThe COVID-19 pandemic posed a unique challenge for summer research programs in 2020, particularly for programs aimed at hands-on experience for younger trainees. The Indiana University Melvin and Bren Simon Comprehensive Cancer Center supports two pipeline programs, which traditionally immerse high school juniors, seniors, and early undergraduate students from underrepresented populations in science in hands-on projects in cancer biology labs. However, due to social distancing policies during the pandemic and reduction of research operations, these students were not physically allowed on campus. Thus, the authors set out to strategically pivot to a wholly virtual curriculum and evaluate the Virtual Summer Research Experience in Cancer outcomes. MethodsThe virtual program included four components: 1. a core science and professional development curriculum led by high school teachers and senior undergraduates; 2. faculty-delivered didactic sessions on cancer science; 3. mentored, virtual research projects with research faculty; and 4. online networking events to encourage vertical mentoring. Outcomes data were measured using an 11-item Research Preparation scale, daily electronic feedback, and structured evaluation and feedback via Zoom weekly. ResultsOutcome data suggested high self-reported satisfaction with the virtual program. Outcome data also revealed the importance of coordination between multiple entities for seamless program implementation. This includes the active recruitment and participation of high school teachers and further investment in information technology capabilities of institutions. ConclusionsFindings reveal a path to educate and train high school and early undergraduate students in cancer research when hands-on, in-person training is not feasible. Virtual research experiences are not only useful to engage students during public health crises but can provide an avenue for cancer centers to expand their cancer education footprints to remotely located schools and universities with limited resources to provide such experiences to their students.

scientific communication and education↗