Keiko Taniguchi-Ponciano

One of the main goals in the treatment of patients with type 2 diabetes (T2D) is the prevention of its complications. Most existing studies have evaluated a limited and isolated number of inflammatory molecules and metabolic factors, and only rarely have clinical and lifestyle variables been jointly considered. The objective of this study is to describe the protocol design for a comprehensive assessment of patients with T2D, considering glycemic control and the development of complications. A cross-sectional, observational, and multicenter study was conducted in three primary care units located in the states of Coahuila, Jalisco, and Veracruz. Glycemic dysregulation will be assessed by measuring glycated hemoglobin, while diabetic nephropathy will be identified through alterations in the glomerular filtration rate. Additionally, clinical and biochemical parameters, lifestyle habits, physical condition, type of treatment, treatment adherence, and nutritional aspects will be evaluated. Furthermore, transcriptomic analysis using RNA-Seq will be performed to identify the genes that best stratify and distinguish patients with glycemic dysregulation, metabolic alterations, and diabetic nephropathy through principal component analysis and ROC curves based on relative RNA expression. To determine clinical factors associated with the link between molecular profiles and glycemic dysregulation, multiple logistic regression models will be developed to identify a pathological molecular signature characteristic of the disease.

Clinically aggressive lactotroph pituitary neuroendocrine tumors (PitNET) are invasive tumors with an unusually rapid growth rate despite maximally tolerated doses of dopamine agonist (DA). We aimed to unravel the molecular heterogeneity of lactotroph PitNET and to identify biomarkers of aggressiveness and resistance to pharmacological treatment. A total of 13 patients harboring DA-resistant lactotroph PitNET were included in this study. Visium Spatial Transcriptomics (ST), whole transcriptome sequencing (WTS), and whole exome sequencing (WES) were performed in tumors from 4 of these patients; WTS and WES was carried out in 5; tumors from two patients underwent ST and WES and tumors from two other patients underwent only ST. Tumors were classified as null or partial responders according to their response to DA treatment. The eight PitNET analyzed by ST exhibited significant intratumoral heterogeneity, with clones showing alterations in PI3K/AKT and lipid metabolism pathways, particularly inositol phosphate, glycerophospholipid, and sphingolipid metabolism. The cell-cell communication analysis showed FGF-FGFR ligand receptor interaction whilst the transcription factors RXRA and CREM showed participation in both groups. A trajectory exploration was performed by including all PitNET together in a single analysis to determine whether there was a tendency or molecular pathway showing a differentiation pattern that would guide the transition from a partially responsive PitNET to a completely unresponsive one. We did not observe any such pattern. All of these findings were corroborated in the cohort of DA-resistant PitNETs in which only bulk WTS and WES were performed. The bulk WTS corroborated lipid metabolism and PI3K-AKT pathway alteration in PitNET, whereas the WES showed only SF3β1 and TP53 variants in one tumor each. Our work suggests that the PI3K/AKT pathway may constitute a molecular target at which to aim therapeutic strategies designed to treat aggressive and DA-resistant lactotroph PitNET.

Cervical cancer (CC) is a public health concern related to the human papillomavirus (HPV) persistent infection. Minichromosome maintenance 2 (MCM2) has been postulated as a surrogate marker for HPV infection. Thymopoietin (TMPO) is a nuclear protein regulated by E2F such as MCM2 or p16. TMPO can give rise to six different isoforms. Herein, both the mRNA and protein levels of TMPO isoforms were analyzed in cervical cells. TMPO expression was selected and analyzed through in silico in several databases from the healthy cervix and cervical lesions. TMPO RNA expression was evaluated in cervical samples and cell lines by RT‐PCR and protein expression by Western‐blot and immunohistochemistry assays. TMPO and MCM2 immunostaining were evaluated in cervical smears. The clinical‐pathological correlation analysis was performed using Kruskal–Wallis or X ² tests. TMPO is overexpressed in 74% of CC cells and all CC cell lines. Moreover, negative immunostaining was observed in normal cervical tissue, compared to strong expression for cervical lesions. Interestingly, TMPO‐α, ‐β, ‐δ, ‐ε, and ‐γ are expressed in all cervical cells and tissues, but a differential expression for α, ‐β, and ‐γ isoforms among the cervical cells was observed as overexpressed when HPV is present. Also, the immunostaining of both MCM2 and TMPO was quite similar, but TMPO expression was more sensitive and specific than MCM2 protein. The present study has revealed that TMPO protein expression could be a potential molecular marker for cervical transformed cells, highlighting the TMPO‐α, ‐β, and ‐γ isoforms as a promising molecular marker of HPV infection.

SUMMARY Familial partial lipodystrophy type 2 is a rare disease, particularly when it is caused by nonclassical gene variants. A high index of suspicion is essential for a timely diagnosis. We present the case of a 32-year-old woman, referred to evaluation of a possible Cushing syndrome, which was clinically and biochemically ruled out. Yet, due to the finding of a rather abnormal fat distribution during physical examination, the diagnosis of lipodystrophy was cogitated. Whole-exome sequencing revealed a missense variant of [...] SUMMARY Familial partial lipodystrophy type 2 is a rare disease, particularly when it is caused by nonclassical gene variants. A high index of suspicion is essential for a timely diagnosis. We present the case of a 32-year-old woman, referred to evaluation of a possible Cushing syndrome, which was clinically and biochemically ruled out. Yet, due to the finding of a rather abnormal fat distribution during physical examination, the diagnosis of lipodystrophy was cogitated. Whole-exome sequencing revealed a missense variant of

Introduction: MODY misdiagnosis remains widespread, existing remarkable variability within genetic variants across populations. While diagnostic tools are based on Caucasian cohorts, Whole Exome Sequencing (WES) studies are needed to identify new genes in non-Caucasians, as up to 77% of patients do not harbor variants of significance in MODY-known genes. In Latino populations, no WES studies have addressed MODY genomic landscape beyond its canonical genes. Methods: We carried out WES in 17 patients with MODY, 17 patients with type 2 diabetes (T2DM) and 17 healthy controls (HC). MODY diagnosis was established according to Exeter criteria (score ≥36%) in subjects with no or minimal insulin requirements. We compared the single nucleotide variant (SNV) landscape across groups. Results: Patients with MODY present a polygenic landscape with allelic variants in canonical and non-canonical genes. Canonical MODY genes used for routine genetic diagnosis showed low discrimination utility, having similar frequencies between MODY, T2DM and HC in the Mexican population. We propose 14 genes with variants that distinguish MODY from T2DM and HC, as we detected variants in genes as MAP2K3, SYT15, TPTE, KCNJ12, PEX5, and OR2A1 in 75-100% of MODY cases while were absent in T2DM and HC. Enrichment analysis revealed involvement in synaptic vesicle trafficking, insulin/IGF pathway-mitogen activated protein kinase kinase/MAPK cascade, and insulin/IGF pathway-protein kinase B/AKT signaling cascade. Discussion: MODY presents a polygenic landscape. Besides improving our understanding of glycemic regulation pathways, the candidate genes could serve as MODY diagnostic biomarkers in Latino populations. Funding: Supported by grant R-2019-785-052 from Instituto Mexicano del Seguro Social.

Pituitary neuroendocrine tumors (PitNET) are known to be variably infiltrated by different immune cells. Nonetheless, their role in pituitary oncogenesis has only begun to be unveiled. The immune microenvironment could determine the biological and clinical behavior of a neoplasm and may have prognostic implications. To evaluate the expression of immune-related genes and to correlate such expression with the presence of infiltrating immune cells in forty-two PitNETs of different lineages, we performed whole transcriptome analysis and RT-qPCR. Deconvolution analysis was carried out to infer the immune cell types present in each tumor and the presence of immune cells was confirmed by immunofluorescence. We found characteristic expression profiles of immune-related genes including those encoding interleukins and chemokines for each tumor lineage. Genes such as IL4-I1, IL-36A, TIRAP, IL-17REL, and CCL5 were upregulated in all PitNETS, whereas IL34, IL20RA, and IL-2RB characterize the NR5A1-, TBX19-, and POU1F1-derived tumors, respectively. Transcriptome deconvolution analysis showed that M2 macrophages, CD4+ T cells, CD8+ T cells, NK cells, and neutrophils can potentially infiltrate PitNET. Furthermore, CD4+ and CD8+ T cells and NK cells infiltration was validated by immunofluorescence. Expression of CCL18, IL-5RA, and HLA-B as well as macrophage tumor infiltration could identify patients who can potentially benefit from treatment with immune checkpoint inhibitors.

Context: The tumor microenvironment (TME) includes diverse cellular components such as mesenchymal stem cells (MSCs) and immune cells, among others. MSC have been isolated from different tumors and they favor tumor cell growth; however, their role in pituitary tumors (PTs) remains unknown. Objective: Herein we report the presence of MSCs in 2 adrenocorticotropin (ACTH)-secreting PTs causing Cushing disease (MCU), 2 nonfunctioning adenomas of gonadotrope differentiation (MNF), and 2 nontumoral pituitary glands (MS). Methods: We have analyzed the transcriptomic profiles by RNA sequencing and compared MSCs in terms of their immunosuppressive effects against lymphoid T-cell and macrophage populations by means of cocultures and flow cytometry. Results: Our transcriptomic analysis revealed molecular differences between MSCs derived from nontumoral pituitaries and MSCs derived from PTs. Two distinct subpopulations of MSC emerged: one displaying immunosuppressive properties and the other with increased proproliferative capabilities, regardless of their origin. MSCs derived from ACTH- and nonfunctioning PTs, but not those derived from nontumoral glands, significantly inhibited the proliferation of activated T cells, favored the generation of regulatory T cells, and promoted M2 macrophage polarization. Such immunosuppressive effects were correlated with an upregulation of programmed death ligand 1 and intracellular expression of macrophage colony-stimulating factor (M-CSF) and interleukin-10. Importantly, MSC derived from ACTH-PTs showed a higher immunosuppressive potential than MSC isolated from nonfunctioning tumors. Conclusion: This study demonstrates the presence of at least 2 MSC subpopulations in the pituitary gland and suggests that immunosuppressive effects of MSCs may have important implications in PT growth.

Clinically non-functioning pituitary adenomas (CNFPAs) are the second most frequent sellar tumor among studies on community-dwelling adults. They are characterized by the absence of hormonal hypersecretion syndrome, and patients present with compressive symptoms, such as a headache and visual field defects. Immunohistochemically, most CNFPAs are of gonadotrope differentiation, with only a few of them being truly null cell adenomas. Although these tumors express receptors for one or more hypothalamic releasing hormones, to what extent this has an impact on the biological and clinical behavior of these neoplasms remains to be defined. In this research, we evaluated the basal and hypothalamic secretagogue-stimulated intracellular calcium mobilization in 13 CNFPAs, trying to correlate this response to the phenotypic features of the patients. Our results indicate that the recurrence of a CNFPA correlates positively with cellular responsiveness, as measured by spontaneous intracellular calcium activity and the ability to respond to multiple hypothalamic secretagogues. We conclude that this finding may be a useful tool for predicting the clinicopathologic behavior of CNFPAs, by testing the variation of cellular responsiveness to hypothalamic secretagogues.