Milestones in Microscopy

Julie In, PhD, a member of the Cellular and Molecular Oncology Research Group, focuses on understanding how intestinal secretory cells respond to acute injury and stress. These secretory changes can be transient and sometimes occur in less than 1% of epithelial cells within the gut. The students in her lab have been trained not only on the ALM equipment but also on cutting edge software for microscopy analysis — such as Huygen’s deconvolution and Leica AIVIA — to understand gut epithelial physiology.

Myranda Thompson, 4^th Year BSGP student, In Lab

Myranda’s project focuses on the secretory response to a potent bacterial toxin, EspP, that is secreted by enterohemorrhagic E. coli. She has been using the Zeiss LSM800 and Olympus/Yokogawa spinning disk confocal microscopes to characterize changes in secretory enteroendocrine cells upon EspP exposure. Her work suggests that acute injury from this bacterial toxin induces enteroendocrine cells to differentiate and increases the amount of hormone that the mucosa secretes as part of a broad immune response.

Cristina Coffman and Haydee Liu, 2^nd Year BSGP students, In Lab

Cristina and Haydee are working together to investigate how heavy metal exposure affects the gut. Cristina is studying changes in the secreted mucus layer and mucus-producing goblet cells and is using the Olympus/Yokogawa spinning disk confocal microscope to quantify the height of the mucus layer. She hypothesizes that heavy metals reduce this layer and contribute to localized inflammation. Haydee is creating transgenic human organoids to examine PROX1. The organoids are three-dimensional models of the human intestine, and PROX1 is a non-canonical transcription factor that may drive secretory cell hyperplasia in response to heavy metal exposure. Haydee will use the Olympus/Yokogawa system’s live-cell imaging capability to define PROX1’s role in enteroendocrine cell differentiation and hyperplasia.

Lito Appell, 1^st Year BSGP student and former UNM U-RISE student, In Lab

Lito is investigating how heavy metal exposure, specifically non-fissile uranium, affects epithelial proliferation and differentiation in the gut. He uses the Olympus/Yokogawa spinning disk confocal microscope to examine miRNAs localization. Lito is using RNAscope fluorescence in situ hybridization of miRNA biogenesis transcripts and specific miRNAs. His study will define how miRNAs regulate secretory differentiation in the gut following acute heavy metal injury.

Nathan Zaidman, PhD, a member of the Cellular and Molecular Oncology Research Group, focuses on the physiological roles of adhesion class G protein-coupled receptors in the kidney. His research aims to identify the function of understudied GPCRs to improve understanding of kidney physiology in health and disease.

Jianxiang (Jason) Xue PhD, postdoctoral fellow, Zaidman Lab

Dr Xue has been using the Leica system to localize mRNAs for Adhesion G-protein-coupled receptors (AGPCRs) to specific types of kidney cells. His imaging demonstrates for the first time that specific AGPCRs are expressed in different kidney cells. His foundational work on the physiological roles of AGPCRs, including Adgrg1, continues in further studies using transgenic mouse models.

Dr Xue’s manuscript, “Identification and localization of adhesion G protein-coupled receptor expression in the murine kidney,” was recently published in the American Journal of Physiology – Renal Physiology. And, his RNAscope image was featured on the cover of AJP Consolidated Print. The image showed Adgrg1 transcripts in relation to Aqp1+ proximal tubule/descending limb cells and Aqp2+ connecting tubule/principal cells.

Ryan Martinez, 2^nd Year BSGP student, Zaidman Lab

Ryan is investigating the Adhesion G-protein-coupled receptors Gpr116 and Adgrf5. His studies focus on how these AGPCRs affect acid secretion from A-type intercalated cells (AICs) in the collecting ducts of mouse kidneys. Ablating Gpr116 from collecting ducts in mice decreases urine pH by allowing V-ATPase proton pumps on the surface of A-type cells to accumulate. Ryan is using the Leica and Spinning Disc confocal microscopes to study the morphological changes in AICs when they secrete acid. He hypothesizes that Gpr116 is necessary to return the activated AIC membrane back to its basal state. Ryan is testing his hypothesis on fixed tissue sections and on isolated AICs from green fluorescent protein (GFP) and tdTomato reporter mice.

Tou Yia Vue, PhD, is a member of the Cellular and Molecular Oncology Research Group. His research focuses on the cellular and molecular mechanisms that underlie the development and function of glial cells (astrocytes and oligodendrocytes) in the Central Nervous System.

Bianca Myers, recent BSGP Graduate, Vue Lab

Bianca extensively used the Leica TCS-SP8 confocal microscope to obtain high-resolution images and tile scans of mouse brain sections. This imaging approach supported her investigation into how the transcription factors ASCL1 and OLIG2 contribute to glioblastoma (GBM) progression. She used transgenic CRISPR-Cas9 induction model of GBM to show that somatic mutations in progenitor cells in the subventricular zone dysregulated ASCL1 and OLIG2.

She identified that ASCL1 influences tumor cell migration and proliferation, two features that drive therapeutic inefficacy. She also demonstrated that the loss of either ASCL1 or OLIG2 reduces tumor growth, while the combined loss of both factors prevents tumor formation entirely. These findings point to molecular targets that may support future therapeutic strategies for addressing GBM. These outcomes represented the final phase of a comprehensive research effort for her dissertation, which resulted in a Nature Communications publication titled “Transcription factors ASCL1 and OLIG2 drive glioblastoma initiation and co-regulate tumor cell type and migration.”

Transcription factors ASCL1 and OLIG2 in glioblastoma.

ASCL1 and OLIG2 are required for tumor formation but inversely regulate different aspects of tumor migration in GBM mouse model. Representative images of tdTOM+ tumors at P30, P60, or terminal stage in control (b–e), Ascl1-CKO (f–i), Olig2-CKO (j–m), and double CKO (n–q, s, t) mice (number of tumors imaged: n=4/genotype for P30 & P60 and n=6/genotype for terminal tumors). Arrows indicate midline and arrowheads mark the distance of migration of tdTOM+ tumor cells on the contralateral corpus callosum (CC). Asterisks demonstrate region imaged for (e, i, m, q, and t). ASCL1 and OLIG2 are highly co-expressed in control tdTOM+ tumor cells, but absent in the single or double CKO tdTOM+ tumors.

Mara Steinkamp, PhD, is a member of the Cellular and Molecular Oncology Research Group and serves as the faculty director of the UNM Comprehensive Cancer Center Animal Models shared resource. She has created several models of ovarian cancer using humanized mice and patient-derived ovarian cancer cells.

Parisa Nikeghbal, recent BSGP Graduate, Steinkamp Lab

Parisa Nikeghbal investigates how tumor-associated macrophages (TAMs) are recruited to ovarian cancer tumors and how they influence those tumors’ responses to treatment. Late-stage ovarian cancer is marked by peritoneal spread and accumulation of malignant ascites containing chemoresistant cancer spheroids and immune cells. TAMs within spheroids and solid tumors establish an immunosuppressive, pro-tumorigenic niche. Using patient-derived organoids co-cultured with human PBMC- or THP-1–derived macrophages, Parisa developed a 3D migration assay to quantify macrophage infiltration using the Zeiss LSM 800 confocal microscope.

Her analyses revealed patient-specific variability in M2 macrophage recruitment, driven in part by tumor-secreted M-CSF, which correlated with increased paclitaxel resistance in 2D cultures, 3D spheroids, organoids, and humanized PDX models. Inhibiting macrophage recruitment with a CSF1R inhibitor reduced tumor regrowth when combined with chemotherapy. These findings demonstrate that individual cytokine profiles modulate TAM infiltration and therapeutic resistance and provide a platform for testing second-line therapies, advancing personalized treatment strategies for OC. Results were published in OncoImmunology.

Macrophage Recruitment by OC Organoids.

Confocal image of fixed PDX3 organoids showing M2 macrophages, derived from PBMCs, migrating and integrating into the organoid structure. DAPI (blue) marks nuclei, E-cadherin (red) highlights epithelial tumor cells, and CD45+ macrophages (green) indicate immune cell infiltration. Scale bar = 50 μm.