bluuebunny/crossref_metadata_2025 huggingface dataset:
Paraneoplastic Hypoglycemia Leading to Insulin Independence in a Patient With Type 1 Diabetes
Authors: Haeri Nami Safai , Mahmud Hussain , Korytkowski Mary T
Published: May 2021

Introduction: Hypoglycemia is a rare paraneoplastic manifestation of several non-islet cell tumors (NICT) that is rarely reported in individuals with diabetes. We report the first case of a patient with type 1 diabetes (T1D) and a recurrent gastrointestinal stromal tumor (GIST) who required discontinuation of all insulin therapy with continuous parenteral glucose infusions to avoid hypoglycemia.

Case: The patient is an underweight (BMI 14.9 kg/m2) 59-year-old female with a 24-year history of T1D treated with insulin pump therapy who was diagnosed with GIST in 2003. Following surgical resection with postoperative adjuvant therapy, she remained tumor free until 2012 when she had disease recurrence unresponsive to several tyrosine kinase inhibitors. In 2014, she reported frequent episodes of documented hypoglycemia despite continued reductions in basal insulin infusion rates. Laboratory testing following several hours of insulin discontinuation revealed undetectable insulin and C-peptide levels, low IGF-1, normal IGF-2, and an IGF-2: IGF-1 ratio of 7.86. Thyroid, kidney and adrenal tests were normal. Initiation of prednisone alone then in combination with octreotide did not abate the hypoglycemia. During a hospitalization for hypoglycemia, exogenous insulin therapy was discontinued and 10% dextrose infusions started. Attempts at weaning the dextrose infusions resulted in hypoglycemia, prompting insertion of a PICC line prior to discharge home with continuous 25% dextrose infusions at 40-60 mg per hour. Discussion: NICT hypoglycemia (NICTH) is characterized by excessive tumor production of IGF-2 or pro-IGF-2. The structure of IGF-2 is similar to insulin, allowing cross reactivity at the insulin receptor. IGF binding proteins (IGFBPs) are responsible for transporting IGF-2 in plasma. Abnormal processing of the gene for IGF-2 results in increased production of the more biologically active pro-IGF-2 that does not form a complex with IGFBP3 and is available to interact with insulin receptors. In patients with normal IGF-2 (chronic kidney disease, poor nutritional status), an IGF-2: IGF-1 ratio >3 can be used to confirm NICTH. IGFBP-3 was not measured in this patient but it is likely this was low due to her poor nutritional status. The most definitive treatment for NICTH is resection of the tumor. Pharmacological management can be considered in patients who are not surgical candidates, but is not always successful as was observed in this patient. Conclusion:This is an unusual case of malignancy associated hypoglycemia in a woman with type 1 diabetes who required discontinuation of all insulin therapy as well as continuous dextrose infusions to achieve euglycemia and briefly maintain an acceptable quality of life over a period of several months.
Presentation of Pheochromocytoma, Papillary Thyroid Cancer and Hyperparathyroidism in Three Family Members
Authors: Ashorobi Damilola , Mandal Kaushik , Liao Huijuan , Kumar Salini Chellappan
Published: May 2021

Introduction: Although rare, one of the most common inherited disorders is multiple endocrine neoplasia. It is an autosomal dominant disorder that predisposes individuals to certain endocrine abnormalities depending on which type. The type 2A is a combination of medullary thyroid cancer, hyperparathyroidism, and pheochromocytoma which have been explained to be due to a mutation in the RET proto-oncogene. This abstract present a case of a patient with hyperparathyroidism whom family members also have pheochromocytoma and papillary thyroid cancer. Case description: A 45 year-old Hispanic male came to the endocrinology clinic complaining of constipation and headache. He has a personal history of non-toxic multinodular goiter and underwent right sided thyroidectomy in 2015 with pathology report showing follicular adenoma. He is currently on thyroid replacement therapy. He is clinically and biochemically euthyroid with TSH of 2.29 IU/ml. Physical examination was unremarkable. His labs were pertinent for calcium 11.5mg/dl, parathyroid 245.7pg/ml, creatinine of 1.5mg/dl. Two years ago, parathyroid was 189.5pg/ml and calcium was 11mg/dl, 1.5 year ago parathyroid level was 235.5pg/ml, calcium was 11.4mg/dl, urine calcium 9.3mg/dl, 24hr urine calcium 286.4mg, calcitonin <2pg/ml, vitamin D 23ng/ml, 1,25 vitamin D 53ng/ml. In 2017, Sestambi scan showed equivocal focus of faint parathyroid activity in the region of the mid to lower left thyroid lobe versus faint residual thyroid activity and in 2019, scan showed no definite parathyroid adenoma. Surgical intervention was recently recommended due to patient’s DEXA scan showing osteoporosis of the femoral neck. The family history of this patient is pertinent for two sisters; one with pheochromocytoma and the other with papillary thyroid cancer. One of the sisters is a 60 years old diagnosed with pheochromocytoma at 51. Her free normetanephrine level was 682pg/ml and total metanephrine was 727pg/ml at time of diagnosis. Her MRI report showed right adrenal mass measuring 3.5x2.8cm. Laparoscopic right adrenalectomy was done and pathology confirmed pheochromocytoma which was RET negative. She still follows up with endocrinology and calcitonin, chromogranin A and plasma metanephrines have been normal. The second sister is now 53 years old diagnosed with papillary thyroid cancer at age 27 and had total thyroidectomy with pathology confirming papillary thyroid cancer. Discussion: Based on the clinical presentation of these family members, the most likely explanation is familial inheritance. This pattern of inheritance cannot be explained by MEN 2A or 2B due to the absence of medullary thyroid cancer. It has also been reported that this unusual presentation could be a variant of MEN 2A.[i] Due to the family history, close follow up is required to monitor for the possible development of other endocrinopathies in the future.
Recurrent Pheochromocytoma, as Component of Multiple Endocrine Neoplasia
Authors: Dominguez Hydelene Beray , Pacheco Elizabeth P , Jimeno Cecilia , Lantion-Ang Franceslina C , Hernandez Edrome , Chiu Harold Henrison Chang
Published: May 2021

Background: Pheochromocytoma is a rare neuroendocrine tumor from the adrenal medulla’s chromaffin cells that secrete catecholamines. The mainstay of treatment is surgery. Although rare, it has a recurrence rate of 6.5-16.5% even after adequate surgical removal with a notable increase in prevalence among genetic syndromes, extra-adrenal tumors, larger tumor size, and younger age of diagnosis. Case: A 23-year-old Filipino male with no known familial disease presented with episodic headache, palpitations, diaphoresis, and resistant hypertension. Two masses in the left adrenal gland, with the larger one measuring 6.0cm x 5.0cm x 3.0cm, were surgically removed. Histologic examination revealed pheochromocytoma. Post-operatively, there was the normalization of urinary metanephrines and the resolution of the signs and symptoms. He was lost to follow-up and returned five years later with an abdominal ultrasound demonstrating a right adrenal mass. He had no accompanying signs and symptoms; the physical examination was unremarkable. On workup, biochemical testing revealed two 24-hour urinary metanephrine levels were markedly elevated, 14.49 and 19.97 (NV:0-1mg/24 hours). Calcitonin: 644 (NV:0-18pg/ml) and carcinoembryonic antigen (CEA): 23.52 (NV:<5ng/ml) were also elevated. Mild hypercalcemia was noted with elevated intact PTH 101.74 (NV:0-65pg/ml). Parathyroid scintigraphy was normal. The neck ultrasound showed multiple bilateral thyroid nodules, with the largest measuring 1.2cmx0.9cm (TIRADS IV). The abdominal CT scan showed two well-defined, homogenous right adrenal masses, with the largest measuring 6.3cm x 5.6cm x 7.4cm. He underwent right adrenalectomy, and histopathology showed pheochromocytoma with Pheochromocytoma of the Adrenal Scale Score (PASS) of 3. One month later, he underwent total thyroidectomy with neck dissection and inferior parathyroid gland resection. Histopathology confirmed medullary thyroid cancer with nodal metastasis and parathyroid adenoma. The patient recovered well without complications. Biochemical tests normalized eight weeks post-surgery. The presence of synchronous recurrent pheochromocytoma, medullary thyroid cancer (MTC), and primary hyperparathyroidism is consistent with multiple endocrine neoplasia 2A (MEN2A) syndrome. Conclusion: Lifelong follow-up is essential in patients treated for pheochromocytoma despite the complete removal of the adrenal masses due to recurrence risk. Recurrence should likewise raise the suspicion of MEN2, a genetic syndrome. In addition, the pheochromocytoma has malignant characteristics, and the MTC has metastases to the cervical lymph nodes. Early detection and prompt intervention are essential for the treatment of the disease.
A Case of Iatrogenic Cushing’s Syndrome
Authors: Del Toro Diez Andrea , Garcia Michelle Marie Mangual , Garcia-Mateo Jose M , Sanchez Ernesto Jose Sola
Published: May 2021

Cushing’s syndrome (CS) is considered a rare disease. The most common cause is the exogenous use of glucocorticoids (GCs), which are often given within a controlled medical setting, but their factitious use is rare. Factitious CS is more common in females, young patients, those with psychiatric disorders, and those with contacts within the medical field. The diagnosis of CS is challenging because some features are non-specific and commonly present in the general population, such as obesity, depression, diabetes, hypertension (HTN), and low bone mineral density (BMD). A high suspicion is warranted. We present the case of a 47-year-old man with HTN, obesity, dyslipidemia, obstructive sleep apnea, and low BMD who complained of increased appetite, significant weight gain, fatigue, sleepiness, muscle weakness, and occasional facial flushing. Medications include Hydrochlorothiazide, Furosemide, Losartan, Atorvastatin, and Teriparatide. Vital signs were normal and body mass index was 41.9 kg/m2. He had a round face, central obesity, and wide purple striae in his abdomen. Dual-energy X-ray absorptiometry scan showed low BMD at spine. Laboratories revealed a glycated hemoglobin of 6.1%, late-night salivary cortisol of <0.03 mcg/dL, 24-hour urine free cortisol of 22.5 mcg/24hr, morning cortisol of 0.01 ug/mL, ACTH 23.5pg/mL, and dehydroepiandrosterone sulfate (DHEA-S) 35 mcg/dL. Our patient persistently denied use of exogenous GCs, but a urine synthetic GC screen disclosed a positive result for dexamethasone; levels at 1.1 mcg/dL. After an exhaustive conversation, our patient confessed to using over-the-counter dexamethasone 4mg to treat occasional muscle aches. ACTH is usually suppressed in factitious CS, but this was not our patient’s case, giving the appearance of ACTH-dependent hypercortisolism. This can lead to unnecessary diagnostic and therapeutic approaches. An unsuppressed ACTH could be due to an unreliable ACTH immunoassay or intermittent, instead of continuous, ingestion of GCs. A suppressed DHEA-S level, as seen in our patient, may provide the clue to exogenous GC use as the cause of CS. Our case is also rare because our patient is male, older, and not related to the medical field. Hypercortisolism must be detected and treated early due to its high morbidity and mortality. Several features may be reversed with treatment. The possibility of hypothalamic-pituitary-adrenal (HPA) axis suppression due to prolonged use of GCs, resulting in adrenal insufficiency (AI) should be considered. The prevalence of GC-induced AI ranges from 14–63%, with the highest risk in those with Cushingoid features and those receiving a dose equivalent to prednisone 20mg daily for more than three weeks. Sudden withdrawal of GCs should be avoided to prevent adrenal crisis. A tapering regimen should be adopted with subsequent biochemical testing of the HPA axis once GCs have been reduced to a physiologic dose.
A Frame-Shift for IGF-1R in Breast Cancer and Metastasis
Authors: Obr Alison E , Lemenze Alexander , Maingrette Krystopher , Ciliento Virginia , Chang Yun-juan , Bulatowicz Joseph J , Wood Teresa L
Published: May 2021

Although, IGF-1R historically has been considered a target for inhibition to treat breast cancer patients, recently, we have identified low levels of IGF-1R are associated with worse overall patient survival and basal-like, triple-negative breast cancer (TNBC). Our findings present a frame-shift for the function of IGF-1R in breast cancer. To elucidate how loss of IGF-1R function impacts aggressive breast cancers, we have integrated the use of human patient data with mouse models where IGF-1R has been deleted or dysregulated. Single cell RNA-sequencing (scRNAseq) analyses from mouse mammary tumors with decreased IGF-1R has revealed expansion of macrophage populations within the tumor consistent with our previous data identifying IGF-1R attenuation promotes an immune evasive tumor microenvironment. Further analysis of the epithelial cell populations identified expansion of the basal and luminal progenitor cells in tumors with reduced IGF-1R. Gene expression alterations in these populations includes enrichment of pathways necessary for cell invasion, cell adhesion, and metastasis. We have further defined how reduced tumor epithelial IGF-1R promotes metastasis by utilizing tail vein injection (TVI). Micrometastatic lesion numbers were similar in animals injected with tumor epithelial cells from control and tumor models with decreased IGF-1R. In contrast to primary tumor-induced metastases, IGF-1R attenuation in dissociated tumor cells decreased metastatic cell proliferation in TVI lungs. Moreover, we observed alterations in expression of several cadherins in tumors with decreased IGF-1R; E-cadherin gene and protein levels are decreased, while P-cadherin gene and protein levels are increased. In vitro cell adhesion assays revealed epithelial cells from tumors with attenuated IGF-1R had reduced adherence to matrix and cell-cell contacts. We have aligned the scRNAseq data from mouse mammary tumors with decreased IGF-1R with human data gene profiles from patient tumors with low IGF-1R and have identified common pathways that are necessary for promoting tumor metastasis including EMT, cell adhesion, and cell cycle. Correlation with the human patient data suggests the mouse models with attenuated IGF-1R reflect similar biology to human patients with low IGF-1R expression. Furthermore, we have uncovered a function for IGF-1R in maintaining proper cell-cell adhesion in the primary tumor supporting the conclusion that dysregulated IGF-1R favors metastatic extravasation of tumor epithelial cells.
A Novel Steroidogenic Factor-1 Antagonist, OR-449, as a Targeted Therapy for Adrenocortical Cancer
Authors: Crowe Paul , Fox Ray , Tao Haiyan , Eastwood Emily , Raheja Neil , Ribeiro Raul , Thacher Scott
Published: May 2021

Adrenocortical carcinoma (ACC) is a rare cancer with an annual incidence of 0.7-2.0 cases per million in the US and EU. Surgical resection combined with adjuvant chemotherapy remains the primary treatment for patients with advanced disease. Although surgery is an option for most patients, oftentimes cases presenting with locally advanced or metastatic disease are not candidates for surgical resection. Furthermore, the non-specific adrenolytic and highly toxic FDA-approved chemotherapy drug, mitotane, is only marginally effective in adult and pediatric ACC. Steroidogenic factor-1 (SF-1 or NR5A1), an orphan nuclear receptor that is essential for the growth and development of the adrenal gland, is highly expressed in adult and pediatric ACC. Moreover, SF- is the major, active transcription factor in ACC (1,2). To address the unmet need for a targeted therapy in ACC, we identified potent small molecule SF-1 antagonists that block SF-1 transcriptional activity through the SF-1 ligand-binding domain (IC50 = 15-20 nM in a CHO cell reporter assay). In short-term dissociated cell cultures established from SJ-ACC3 (3), a pediatric ACC patient-derived tumor xenograft (PDX), OR-449 and a related SF-1 antagonist, OR-907S, blocked DNA synthesis as measured by inhibition of EdU incorporation in SF-1+ cells (IC50 = 500-600 nM, >80% efficacy at 10 μM) whereas OR-907R, the 100-fold less potent enantiomer of OR-907S, is nearly inactive. OR-449, which has >20% oral bioavailability and a long plasma half-life (> 9 h) in mouse, rat and dog, inhibited growth of SJ-ACC3 tumors grown in immunocompromised mice following daily oral dosing (30 mg/kg) for 4 weeks. SF-1-responsive genes, both down- and up-regulated, identified by RNAseq (by comparison of OR-907S and OR-907R in SJ-ACC3 dissociated cell culture), also responded to OR-449 in SJ-ACC3 xenografts following 7 days of dosing, indicating transcriptional regulation of SF-1 by OR-449. Importantly, in an exploratory 2-week mouse safety study, OR-449 showed no adverse effects when dosed up to 100 mg/kg. Taken together, these findings suggest that SF-1 antagonists could provide a safe and effective targeted therapy for ACC. References: (1) Mohan, et al., Curr. Opin. Endocrinol. Metab. Res., 2019; 8:72; (2) Corces, et al., Science, 2018; 362:eaav1898; (3) Pinto, et al., Clin. Cancer. Res., 2013; 19:1740.
Amphiregulin-EGFR Autocrine Signaling Mediates Neutrophil Elastase-Triggered Prostate Cancer Progression
Authors: Xiao Zhiguang , Hammes Stephen R
Published: May 2021

Neutrophil elastase (NE) is a serine protease stored in neutrophil azurophilic granules. Growing evidence indicates that NE is intimately involved in the activities of proinflammatory cytokines / chemokines, growth factors, and cell surface receptors. These molecular regulations can modulate innate immune responses as well as directly promote cancer cell outgrowth. To date, however, little is known regarding the molecular mechanisms underlying the stimulatory properties of NE in cancer cells. Here we examine NE effects on prostate cells, demonstrating that NE triggers proliferative signals and cell migration in six prostate cell lines representing the spectrum of prostate cell malignancy, including normal prostatic epithelium, benign prostatic hypertrophy, and metastatic prostate cancer. Using ERK activation as a read-out, we show that NE promotes ERK phosphorylation in a dose dependent manner, and time course study further reveal a sustained ERK activation upon NE treatment. Western blot evaluation demonstrates strong EGFR expression in cell lines derived from normal and benign prostatic gland, and preferential expression in hormone resistant versus hormone responsive cells. In agreement with EGFR-dependent mitogenic signaling, activation of ERK is abrogated by siRNA-mediated EGFR knockdown, as well as by pretreatment of cells with irreversible EGFR inhibitor AG1478. Importantly, NE evokes cancer cell migration at a lower range of NE concentrations relative to nonneoplastic cells. In prostate cells, from a total of seven EGFR ligands, amphiregulin (AREG) is predominantly expressed, and the addition of NE results in the release of AREG. Moreover, AREG gene silencing by siRNA or inhibition of AREG biological activity by neutralizing antibody, prevents NE-induced ERK phosphorylation and cell migration. Together, our study reveals a distinct and essential role of AREG-EGFR signaling axis in NE-triggered prostatic cellular response.
FOXO1 Mitigation of FOXL2C143W/SMAD3 Transcriptomic Landscape in a Model of Granulosa Cell Tumor
Authors: Secchi Christian , Benaglio Paola , Mulas Francesca , Belli Martina , Stupack Dwayne , Shimasaki Shunichi
Published: May 2021

Background: Adult granulosa cell tumor (aGCT) is a rare type of stromal cell malignant cancer of the ovary. Postmenopausal genital bleeding is the main aGCT clinical sign which is attributed to estrogen excess driven by CYP19 upregulation. Typically, aGCTs that are diagnosed at an initial stage can be treated with surgery. However, recurrences are mostly fatal1. Current studies are focused on finding new molecular markers and targets that aim to treat the aGCTs recurrence. Between 95-97% of aGCTs harbor a somatic mutation in the FOXL2 gene, Cys134Trp (c.402C<G)2. A TGF-β pathway protein, SMAD3, was identified as an essential partner in FOXL2C134W transcriptional activity driving CYP19 upregulation3. Recently, the antitumoral FOXO1 gene has been recognized as a potential target for suppressing the FOXL2C134W pathogenic action4. Aim: The objective of this study was to examine whether FOXO1 upregulation affects the FOXL2C143W/SMAD3 transcriptomic landscape. Methods: RNA-seq analysis was performed comparing the effect of FOXL2WT/SMAD3 and FOXL2C143W/SMAD3 overexpression in presence of FOXO1 by transfection of an established human GC line (HGrC1). RNA-seq libraries were prepared using the illumina TrueSeq and sequenced using an illumina HiSeq Platform4000. To quantify transcript abundance for each sample we used salmon (1.1.0) with default parameters, using indexes from hg38. Data was subsequently imported in R using the tximport package and processed with the DESeq2 package. Results: RNA-seq data show that FOXL2C143W/SMAD3 significantly drives 717 genes compared with the WT and enabled us to identify targets (TGFB2, SMARCA4, HSPG2, MKI67, NFKBIA) and neoplastic pathways directly associated with the mutant. To provide evidence that the differences in gene expression were attributed to a direct consequence of FOXL2 binding, we annotated gene promoters with previously published FOXL2 ChIP-seq analysis. The majority (73-40%) of the differential expressed genes (DEGs) between FOXL2C134W and FOXL2WT had a FOXL2 binding site at their promoters, which was a significantly higher proportion than in non-DEGs (Fisher’s exact test, murine: p= 7.9x10-157; human, p= 9.9x10-39). Surprisingly, the number of DEGs between FOXL2C134W + FOXO1 and FOXL2WT was much lower (230) with respect to the number of DEGs between FOXL2C134W and FOXL2WT (717, of which 130 in common; linear regression slope ß = 0 .58), suggesting that the effect of FOXL2C134W compared with FOXL2WT is moderated by the addition of FOXO1. Conclusions: Our transcriptomic study provides the first evidence that FOXO1 can efficiently mitigate 40% of the altered genome-wide effect specifically related to FOXL2C134W in a model of human aGCT.1 Farkkila, A. et al. Ann Med (2017). 2 Jamieson, S. & Fuller, P. J. Endocr Rev (2012). 3 Belli, M. et al. Endocrinology (2018). 4 Belli, M et al. J Endocr Soc (2019).
Heterogeneous Nuclear Ribonucleoprotein K Is Involved in the Estrogen-Signaling Pathway
Authors: Iwabuchi Erina , Miki Yasuhiro , Ito Kiyoshi , Ishida Takanori , Sasano Hironobu
Published: May 2021

Heterogeneous nuclear ribonucleoprotein K (hnRNPK) has been found in the nucleus, cytoplasm, and mitochondria. It is implicated in chromatin remodeling, transcription, splicing, and translation processes. Although hnRNPK has reportedly been associated with poor prognosis in colon cancer patients, it is beneficial in gastric cancer as it inhibits cancer cell proliferation. Expression of hnRNPK in ER (Estrogen receptor) -positive/PR (Progesterone receptor) -positive breast cancer was higher than other subtypes; however, the biological functions of hnRNPK in the ER-mediated signaling pathway have not been identified. In this study, we investigated the functions of hnRNPK in the estrogen-signaling pathway. We initially evaluated hnRNPK expression upon treatment with estradiol (E2) and ICI 182,780 in ERα-positive breast cancer cell line MCF-7. This initial evaluation revealed that expression of hnRNPK was increased by E2 treatment but decreased by ICI 182,780 treatment. We further evaluated the effects of estrogen-signaling pathway in hnRNPK knockdown MCF-7 cells using siRNA, which revealed that hnRNPK knockdown decreased ERα expressions and ERα target gene TFF1 by E2 treatment. In addition, we examined the interaction between hnRNPK and ERα because hnRNPK has been reported to interact with several other proteins. These interactions were detected using immunoprecipitation and proximity ligation assay. We then immunolocalized hnRNPK in breast cancer and endometrial cancer. hnRNPK expression was significantly higher in ERα-positive cancer cells in both breast and endometrial cancers. In contrast, hnRNPK expression was significantly lower in Ki-67-positive breast cancer while being significantly higher in Ki-67-positive endometrial cancer. hnRNPK has been found to function differently, depending on the type of cancer (breast or endometrial) that it is expressed in. However, further studies are required to clarify the clinical significance of hnRNPK in breast and endometrial cancer patients.
In-Vitro Effects of Active Vitamin-D3 and Vitamin-A on Human Melanoma (BLM and1205Lu) Cell Growth
Authors: Ramaraj Pandurangan
Published: May 2021

Though UV ray in the Sun light is essential for vitamin-D3 formation, yet UV rays can lead to skin cancer. Lack of vitamin-D3 also can lead to cancer. Hence, we decided to study the effect of vitamin-D3 on human melanoma cell models. Our aims were to study the in-vitro effects of vitamin-D3 and vitamin-A on human melanoma (BLM, 1205Lu) cell growth, as vitamins D3 and A are important for a healthy skin. Initially dose-curve (100 nM to 100 μM concentration) study was carried out with vitamin-D3 and vitamin-A (retinoic acid) on BLM cells to determine the optimal concentrations of vitamins-D3 and A for co-incubation with progesterone (50 μM) and RU-486 (50 μM). Supernatants from the treated cells were subjected to Elisarray. Vitamin-D3 and vitamin-A showed a dose-dependent decrease in cell growth. Based on the dose-curve, it was decided to use 25 μM (at 57% cell growth) of vitamin-D3 and 50 μM (at 55% cell growth) of vitamin-A for co-incubation studies. Co-incubation of vitamin-D3 with vitamin-A showed an additive effect on the decrease of BLM cell growth (20%). Similarly co-incubation of vitamin-D3 with progesterone (33% cell growth) and with RU-486 (28% cell growth) as well as co-incubation of vitamin-A with progesterone (31% cell growth) and RU-486 (18% cell growth) showed an additive effect on the decrease of BLM cell growth. Based on the BLM co-incubation studies, we decided to repeat the studies on 1205Lu cells. So, 25 μM (at 34% cell growth) of vitamin-D3 and 50 μM (at 44% cell growth) of vitamin-A were used for co-incubation studies. Co-incubation of vitamin-D3 with vitamin-A showed an additive effect on the decrease of cell growth (22%). Though co-incubation of vitamin-D3 with progesterone did not show any difference in cell growth (51%), yet co-incubation with RU-486 showed a decrease in 1205Lu cell growth (21%). Similarly co-incubation of vitamin-A with progesterone (25% cell growth) and with RU-486 (23% cell growth) showed a synergistic effect on the decrease of 1205Lu cell growth. Conclusion: These studies suggested that combination of vitamins and steroids might be effective in decreasing melanoma cell growth. Hence, various combination of vitamins and steroids will be tested for their effect on melanoma cell growth in vitro in order to develop a combo drug treatment for melanoma.
Macrophage-Derived Endotrophin Supports Tumor Migration Potentials in Thyroid Caner
Authors: Shin Hyo Shik , Kim Hana , Song Young Shin , Sun Hyun Jin , Whang Young Mi , Park Jiyoung , Park Do Joon , Park Young Joo , Cho Sun Wook
Published: May 2021

Endotrophin (ETP), a cleaved fragment of the C5 domain of the Type VI collagen α3 (Col6α3), has been shown to play pro-tumorigenic roles in breast and liver cancers. However, the ETP actions in tumor microenvironment (TME) is still undetermined. This study aimed to investigate the role and the mechanism of ETP in macrophage-enriched thyroid cancer TMEs. First, the expression of ETP on various human thyroid tissues was studied. Immunohistochemical staining showed that the ETP was expressed in papillary (PTC) or anaplastic (ATC) thyroid cancer tissues but not in benign adenoma or autoimmune thyroiditis. Among 146 PTCs, tumors were divided into two groups according to their ETP expression, ETPlow and ETPHigh. Tumor size was bigger and LN metastasis rates were higher in ETPHigh than ETPlow group. Interestingly, the ETP expression showed positive correlations with the CD163-positive macrophages in thyroid tumors. To clarify the origin of ETP in TME of thyroid cancer, xenograft tumor from FRO (ATC) cells was co-stained with anti-ETP and anti-F4/80 antibodies. Immunofluorescent staining showed that the expression of ETP and F4/80-postive macrophages were co-localized in the peritumoral area. Additionally, human monocytic THP-1 cells were treated with conditioned media (CM) from FRO or BCPAP cells (designated FRO-CM or BCPAP-CM) and demonstrated that ETP expression was more up-regulated in the FRO-CM or BCPAP-CM treated group than in the control group. Collectively, ETP produced from macrophages in thyroid cancer microenvironments. Next, the pro-tumorigenic functions of ETP has been studied. CMs from co-cultures of FRO or BCPAP cells with THP-1 cells were harvested and treated to FRO or BCPAP cells. Treatment of CMs from co-cultures increased cell migration potentials than that of the single cell alone, and these effects were reduced by anti-ETP neutralizing antibody, indicating that ETPs play essential role in macrophage-induced tumor cell migration potentials. Moreover, MMP-9 and MMP-14, the candidates of Col6α3 protease to produce ETP were increased in THP-1 cells by treatment of CM. Additionally, treatment of CMs from MMP-9 inhibitor-treated co-cultures of FRO and THP-1 cells decreased cancer cells migration potentials than the control CMs. Finally, the RNA-seq dataset of human thyroid cancer showed that higher expressions of ETP positively correlated with macrophage-related genes such as CD163 and MMPs including MMP-9 and MMP-14. In conclusion, macrophage contributes ETP productions by modulating MMP expressions and ETP supports pro-metastatic potentials of human thyroid cancer cells in TMEs. Thus ETP can be a good therapeutic target for macrophage-enriched advanced thyroid cancers.
Prostate Cancer Stimulation by a Novel Liver X Receptor (LXRa)-Estrogen Related Receptor (ERRa) Axis
Authors: Song Chung Seog , Chatterjee Bandana
Published: May 2021

Background: Liver X Receptors (LXRα, β) are oxysterol sensing nuclear receptors that regulate lipogenesis, cholesterol homeostasis and immune cell function. While oxysterols are agonist ligands of LXRs, sulfated oxysterols, catalytically produced by the SULT2B1b sulfotransferase, are LXR-inert. Increased SULT2B1b expression leads to attenuation of LXR signaling. In a previous report, we showed that SULT2B1b is undetectable in clinical samples of castration-resistant prostate cancer (CRPC), and its level is significantly reduced in a subset of primary prostate cancer. In cell models, genetic ablation of SULT2B1b exacerbated aggressive traits of CRPC, evident from EMT-like activation, enhanced invasion, faster xenograft tumor growth and reduced cell adhesiveness and stiffness in single-cell atomic force microscopy analysis. AKR1C3, which promotes androgen biosynthesis and shows elevated expression in advanced prostate cancer, is markedly upregulated in SULT2B-depleted cells. Elevated AKR1C3 leads to activation of the ERK1/2 Map kinase survival signal in CRPC cells. Results: We report here that AKR1C3 upregulation is a consequence of enhanced LXRα signaling in SULT2B1b-deficient cells, since the upregulation was abolished in multiple cell models when LXRα was silenced by siRNAs or inactivated by the small molecule inhibitor SR9243, which is an LXRs-selective inverse agonist. Conversely, LXR agonism induced by an oxysterol, or by the synthetic ligand GW3965, resulted in elevated AKR1C3 expression. Consistent with a recent report that the nuclear receptor ERRα is a positive regulator of AKR1C3, we found that ERRα ablation prevented AKR1C3 upregulation in SULT2B1b-deficient cells. Notably, LXRα inactivation caused marked reduction of ERRα, indicating that ERRα functions downstream of LXRα to induce AKR1C3 and ERK1/2. Dependence of ERRα and AKR1C3 expression on LXRα was observed in both androgen receptor (AR)-positive and AR-negative CRPC cells. Elevated ERRα in prostate cancer is known to be associated with a poor disease outcome. This association may be in part due to ERRα activation by cholesterol, which is the endogenous agonist ligand for ERRα (Cell Metab 23: 479, 2016), and high cholesterol is a risk factor for aggressive prostate cancer. Furthermore, statins, which inhibit cholesterol biosynthesis, are beneficial to CRPC patients with elevated blood cholesterol. We identified two cholesterol-responsive ERRα-binding sites in the far upstream region of the AKR1C3 promoter. This result confirms that ERRα plays a direct role in the transcriptional upregulation of AKR1C3. Significance: Our study establishes a novel LXRα→ERRα→AKR1C3→ERK1/2 survival axis that is activated in CRPC cells under SULT2B1b deficiency. The LXRα→ERRα regulatory axis may be exploited for developing novel therapeutics against AR-positive and AR-negative CRPC.
Small Molecule Modulation of MEMO1 Protein-Protein Interactions
Authors: Pollock Julie A , Labrecque Courtney L , Hilton Cassidy N , Airas Justin , Blake Alexis , Rubenstein Kristen J , Parish Carol A
Published: May 2021

MEMO1 (mediator of ErbB2-driven cell motility) is upregulated in breast tumors and has been correlated with poor prognosis in patients. As a scaffolding protein that binds to phosphorylated-tyrosine residues on receptors such as estrogen receptor and ErbB2, MEMO1 levels can influence phosphorylation cascades. Using our previously developed fluorescence polarization assay, we have identified small molecules with the ability to disrupt the interactions of MEMO1. We have performed limited structure-activity-relationship studies and computational analyses to investigate the molecular requirements for MEMO1 inhibition. The most promising compounds exhibit slowed migration of breast cancer cell lines (T47D and SKBR3) in a wound-healing assay emulating results obtained from the knockdown of MEMO1 protein. To our knowledge, these are the first small molecules targeting the MEMO1 protein-protein interface and therefore, will be invaluable tools for the investigation of the role of the MEMO1 in breast cancer and other biological contexts.
Targeting Glycogen Metabolism as a Novel Therapeutic Approach in Anaplastic Thyroid Cancer
Authors: Davidson Cole , Tomczak Jennifer , Amiel Eyal , Carr Frances
Published: May 2021

Effective treatment options for well-differentiated papillary (PTC) and follicular (FTC) thyroid cancers afford positive patient prognoses. The absence of effective interventions for the stem-like, dedifferentiated anaplastic thyroid cancer (ATC) results in poor patient outcomes with a mortality rate higher than all other endocrine cancers combined (1). While receptor tyrosine kinase inhibitors such as sorafenib can extend ATC patient survival, drug resistance and tumor reoccurrence often develop (2). Therefore, there is a critical need for more effective targeted therapies for ATC. Although the cell signaling landscape of ATC tumors is well described, very little is known about tumorigenic adaptations in ATC cellular metabolism. Tumors exhibit an increased consumption of glucose compared to normal tissues to fuel tumor progression. Some cancers meet this high glucose requirement by storing and breaking down glycogen. In our studies here, we show for the first time that normal thyroid, PTC, FTC, and ATC cells express genes necessary for glycogen metabolism. We confirm these observations in patient samples in normal thyroid and thyroid cancer patient samples via immunofluorescence in tissue microarrays. Furthermore, we detect intracellular glycogen stores in cell lines representing normal thyroid, PTC, FTC, and ATC cells. Importantly, we demonstrate that glycogen phosphorylase inhibitors result in accumulation of intracellular glycogen and induce subsequent apoptosis in ATC cells. We further show that glycogen phosphorylase inhibitors synergize with kinase inhibitors such as sorafenib and buparlisib to decrease ATC cell viability. Our work establishes glycogen metabolism as a novel metabolic process in thyroid cells that is associated with thyroid cancer dedifferentiation and provides insight to the effectiveness of inhibiting glycogen metabolism as a therapeutic strategy in ATC.

References: 1. Siegel, R.L., Miller, K.D. and Jemal, A. (2017), Cancer statistics, 2017. CA: A Cancer Journal for Clinicians, 67: 7-30. doi:10.3322/caac.21387

2. Saini S, Tulla K, Maker AV, Burman KD, Prabhakar BS. Therapeutic Advances in Anaplastic Thyroid Cancer: A Current Perspective. Mol Cancer. 2018;17(1):154. doi:10.1186/s12943-018-0903-0
A Rare Cause of Recurrent Hypoglycemia - The “Big-IGF-II” Factor
Authors: Syeda Asma , Mohammed Turab , Padmanabhan Premkumar
Published: May 2021

Introduction: Hormone secreting tumors are a rare cause of hypoglycemia. Here we present an interesting case of paraneoplastic phenomenon resulting in a non-islet cell tumor causing hypoglycemia (NICTH) from a solitary fibrous tumor (SFT). Clinical Case: An 83-year-old female with a history of hypertension presented with recurrent episodes of lightheadedness and falls for one month. Review of systems was negative except for insignificant weight loss. Her vital signs and physical examination were unremarkable. Only significant finding on initial blood work was a blood glucose level of 30 mg/dL (n: >70mg/dL). Additional work-up of hypoglycemia revealed a low serum free insulin level 0.6 IU/mL (n:1.5 - 14.9 IU/mL), low proinsulin <4 pmol/L (n: <18.8 pmol/L), low C-peptide level 0.14 ng/mL (n: 0.80-3.85 ng/mL), low serum insulin-like growth factor-I (IGF-I) 33 ng/ml (n:34-246 ng/ml) and elevated IGF-II levels 370 ng/mL (n:47-350 ng/mL). Blood sulfonylurea and insulin antibodies screen were negative, ACTH and morning cortisol levels were within normal limits. A CT scan of chest, abdomen and pelvis for neoplastic workup demonstrated a large heterogeneous mass within the lower lobe of the right lung. A CT-guided biopsy disclosed spindle cell neoplasm, positive for signal transducer and activator of transcription-6 (STAT6) and CD34. Reverse transcription-polymerase chain reaction (RT-PCR) confirmed the NGFI-A binding protein 2 (NAB2) and STAT6 fusion which is specific for solitary fibrous tumor (SFT). She underwent surgical resection of the thoracic mass which resulted in complete resolution of her symptoms. Clinical Lessons: NICTH is a paraneoplastic syndrome associated with various benign and malignant tumors including SFT, hemangiopericytoma and metastatic hepatocellular carcinoma to name a few. Our case is an example of NICTH associated with SFT called Doege-Potter syndrome. The prevalence of hypoglycemia is seen in less than 5% of patients with SFT. It is often secondary to ectopic overexpression of unprocessed precursor ‘big’ IGF-II molecule, which binds to IGF-I receptor, resulting in tumorigenesis. An IGF-II/IGF-1 ratio is a sensitive marker and a value of more than 10 suggests excess production of IGF-II and is likely the cause of NICTH. Physicians should be cognizant of NICTH, especially in non-diabetic, otherwise healthy individuals in whom preliminary workup for hypoglycemia is negative and should look to identify any potential tumor as the likely cause. Surgical treatment is often the only curative option. Frequent meals, dextrose infusion, glucagon and growth hormone are bridging modalities until definitive surgical treatment.
Late Onset of Abiraterone Severe Hypokalemia Due to Unsuspected ACTH-Secreting NEC
Authors: Toral Fernando Pazos , Duran Ignacio , Schettini Isabel Alvarez
Published: May 2021

Clinical Case: 68-year-old male, Active smoker (30-40 packs/year), and moderate alcohol drinker. An increase in urinary frequency, nocturia, and a PSA of 116 µg/L led to the diagnosis of disseminated castration-sensitive prostate cancer (CSPC), advanced Gleason score (3 + 4 pT2), with bone and retroperitoneal metastases (CT). By 02/2018, he started androgen deprivation therapy (ADT) with Abiraterone (AB) 1000 mg, prednisone, and LHRH agonist. 15 months later, PSA levels decreased to 0.75 ng/ml without side effects and normal K+(3.7 mEq/L), when a CT scan revealed both liver and bone metastasis. During the progression study, two months later, the patient was admitted to the hospital for severe hypoK+ (1.7 mEq / L), normal renal function and metabolic alkalosis. Although abiraterone was discontinued, up to 460 mEq iv per day of K+ and spironolactone were required to maintain serum K+ above 2.5. 3 days later, the hormonal study revealed TSH 1.13 mU /L, ACTH 162 pmol/L, cortisol 258 nmol/L, aldosterone 105 pmol/L, renin (protein) 0.7 µU/mL and deoxycorticosterone 507 pmol/L. 11 days later, plasma cortisol was 967 nmol/L, ACTH 132pmol/L, and cortisoluria 1456 nmol/d. The suppression test with 1 mg of DXM for cortisol was 1162,5 nmol/L. DHEAS was < 407 nmol/L. Liver biopsy showed a small cell neuroendocrine carcinoma (NEC), chromogranin (+), synaptophysin (+), CD56 (+), TTF-1 (+), PSA (-), Ki-67 90% and the granular cytoplasmic ACTH (+). The octreotide scan (+) revealed pathological uptake in L4, multiple uptakes in the liver, and in the axial skeleton. Treatment with Carboplatin plus Paclitaxel for NEC was started, completing 3 weekly doses with good tolerance and clinical benefit, but with the persistence of severe hypoK+ (2.5 mEq/L). Treatment with lanreotide 120 mg every 4 weeks was added, following a feeling of clinical improvement, the disappearance of edema, asthenia, and normalization of plasma K+ (3,4mEq/L). The patient died two months later from respiratory sepsis. Discussion: Several clinical trials have demonstrated that the combination of AB plus ADT prolongs overall survival in DCSPC. The CYP17 inhibition by AB increases ACTH leading to early secondary mineralocorticoid excess with hypokalemia and hypertension while the cortisol levels remain normal or low. Cortisol serum levels increased after AB was discontinued, whereas aldosterone serum levels remained low due to K+ regulatory feedback. The hormonal profile and the pathological and radiological studies revealed an ACTH-producing small cell NEC. In this patient, the previous treatment with BA has masked the clinical and hormonal profile of an ectopic Cushing syndrome. Therefore, Cyp17 inhibitors can mask adrenal or extra-adrenal processes characterized by alterations in steroid metabolism. This case has suggested a more thorough assessment of the adrenal hormonal profile including ACTH during BA treatment.
Salivary Estrogens Androgens and Aromatase Inhibitors by LC-MS/MS in Breast Cancer Patients Treated With Testosterone Pellets Combined With Aromatase Inhibitors
Authors: Zava David T , Kimball David W , Glaser Rebecca L
Published: May 2021

Introduction: High dose testosterone (T) has been used for treating hormone-sensitive breast cancers for many years. However, a drawback to T therapy is its propensity to convert to estradiol (E2) via aromatase, which can override the growth inhibitory effects of T and stimulate estrogen sensitive tumors. Aromatase is higher in some women than others, particularly those with active tumors, truncal obesity, and inflammatory conditions contributing to and caused by cancer. Oral aromatase inhibitors (AI) have been used to prevent conversion of T to E2. While this effectively reduces E2 burden, systemic oral AI require higher dosing, which often leads to severe side effects. As an alternative to oral AI therapy, researchers have found that T combined with a much lower dose of AI in a solitary pellet implant placed in the subcutaneous (SC) tissue, or alternatively, into the breast adjacent to a primary tumor, is effective in reducing tumor burden and maintaining a low systemic level of E2, while reducing the adverse side effects of very low E2. Study Design: In this case study we report on the use of an LC-MS/MS method to monitor salivary levels of E2, Estrone (E1), T, and the AIs Letrozole (LET) and Anastrozole (ANZ) following T + AI therapy in a breast cancer patient (intolerant of oral LET) with active (measurable) tumor in the breast and metastatic disease. Steroids and AIs were measured in saliva at baseline, 1 week, and 4 weeks after insertion of subcutaneous pellets containing ‘60 mg T + 4 mg ANZ’ and ‘60 mg T + 6 mg LET’. Results: LC-MS/MS (range values-pg/ml) for postmenopausal women, and baseline, week 1 and week 4 post treatment values for each steroid and AI were: E2: (range 0.3-0.9), 0.4, < 0.3, < 0.3; E1: (range 0.9-3.1), 1.0, <0.4, <0.4; T: (range 7-22), 6, 96, 48; ANZ: (range <4), < 4, 2063, 24; LET: (range <4), < 4, 744, 175. Self-reported (Pre/Post Therapy) estrogen deficiency symptoms such as hot flashes, night sweats, vaginal dryness, joint pain, and sleep disturbances were significantly improved post T+AI therapy. In addition, the intramammary tumor decreased in size > 95% by month 5. Summary: These results show that low-dose SC AI therapy (4-6 mg/2-3 months = 0.1 mg/day) with T (120 mg/2-3 months = 1-2 mg/day) increases T to supra-physiological levels, prevents T metabolism to estrogens E2 and E1, reduces estrogen deficiency symptoms, and has beneficial effects on tumor growth inhibition. Simultaneous testing of sex-steroids E2, E1, T, and aromatase inhibitors LET and ANZ by LC-MS/MS provides a convenient means to monitor the bioavailable levels of these analytes and adjust them as necessary to optimize therapeutic efficacy and reduce adverse side effects.
Think Outside the Thorax: Doege-Potter Syndrome Presenting as a Retroperitoneal Mass
Authors: Gray Cater Taylor Lauren , Symczyk Oksana , Haider Adnan
Published: May 2021

Introduction: Non-islet cell tumor hypoglycemia (NICTH) is a rare paraneoplastic syndrome, occurring less commonly than insulinoma. These tumors produce high molecular weight insulin-like growth factor-2 or “big IGF-2”, resulting in hypoglycemia: big IGF-2 decreases glycogenolysis, gluconeogenesis, and glucagon release as well as increases glucose uptake by adipocytes and skeletal muscle. NICTH specifically associated with a solitary fibrous tumor (SFT) is known as Doege-Potter syndrome. Doege-Potter syndrome is seen most with thoracic SFT’s. This case study details the diagnosis and management of Doege-Potter syndrome due to retroperitoneal SFT.

Case Description: Our patient is a 68-year-old, white female who initially presented to an outside facility with a 3-week history of episodic neuroglycopenic symptoms temporarily relieved with eating. She reported night sweats but no weight change or fever. Medical history was notable only for hypertension and GERD. She had no personal or family history of diabetes. She was transferred to our facility for further evaluation of an 18.6 cm multilobular mass seen just inferior to the liver on abdominal CT. Whipple’s triad was confirmed with venous glucose sampling while inpatient. C-peptide, free/total insulin, and pro-insulin were collected during permissive hypoglycemia and were low. Morning cortisol and TSH were unremarkable. Additionally, sulfonylurea screen and insulin antibodies were negative. Ultrasound-guided biopsy of the lesion was positive for CD34 and STAT6, consistent with SFT. Refractory hypoglycemia was treated with D5W infusion until surgical resection of the retroperitoneal mass, after which, she had complete resolution of her hypoglycemia.

Discussion: This patient’s serum IGF-2 was within normal limits, consistent with reports of feedback inhibition exerted by increased levels of big-IGF-2. Therefore, the IGF-2:IGF-1 ratio is used to determine increased serum levels of big IGF-2, but this test is not widely available. SFT’s can recur within months to years of resection, necessitating surveillance. Recurrence is also more common with extrathoracic tumors. Surgical pathology noted tumor dimensions of 22 cm and 2270 g; mitotic rate was 1 per 10 hpf. Features typical of malignancy include large size (>15 cm) and mitotic rate >4 mitoses per hpf. This patient’s follow-up CT abdomen at 6 weeks did not show any findings to suggest recurrent or metastatic disease.