Welcome to the Resource Library at wrenlaboratories.com. Navigate our different categories of resources below on the left. Next to the different resources, you will find the latest articles about the world of neuroendocrine disease research and testing.

The Identification of Gut Neuroendocrine Tumor Disease by Multiple Synchronous Transcript Analysis in Blood

Irvin M. Modlin, Ignat Drozdov, Mark Kidd

We developed a multi-transcript molecular signature for PCR-based blood analysis. NEN transcripts were identified by computational analysis of 3 microarray datasets and examined in 130 blood samples. Gene-based classifiers detected NENs in independent sets with high sensitivity (85-98%), specificity (93-97%), PPV (95-96%) and NPV (87-98%). A panel of 51 marker genes differentiates NENs from controls with a high PPV and NPV (>90%), identifies pancreatic and gastrointestinal NENs with similar efficacy, and confirms GEP-NENs when CgA levels are low. The panel is significantly more accurate than the CgA assay. This reflects its utility to identify multiple diverse biological components of NENs. Application of this sensitive and specific PCR-based blood test to NENs will allow accurate detection of disease, and potentially define disease progress enabling monitoring of treatment efficacy.

Gut Neuroendocrine Tumor Blood qPCR Fingerprint Assay: Characteristics and Reproducibility

Irvin M. Modlin, Ignat Drozdov, Mark Kidd

We have developed a PCR-based tool that measures a 51-gene panel for identification of gastro- enteropancreatic (GEP) neuroendocrine neoplasms (NENs) in peripheral blood. This manuscript assesses the robustness (performance metrics) of this tool with a specific focus on the effects of individual parameters including collection, storage, acid suppressive medica-tion (proton pump inhibitor (PPI)), age, sex, race and food on accuracy.

Blood and Tissue NET Gene Cluster Analysis correlate, define Hallmarks and Predict Disease Status

Irvin M. Modlin, Ignat Drozdov, Mark Kidd

We evaluated whether blood measurements correlated with tumor tissue transcript analysis. Regulatory network analysis, linear modeling, principal component analysis (PCA), and receiver operating characteristic analyses were used to delineate neoplasia ‘hallmarks’ and assess NETest predictive utility. Our results demonstrated: matched blood/tumor expression was highly significant (R2>0.7, p<0.0001). Integration of algorithm-derived scores and gene expression had an AUC of 0.92+0.02 for differentiating progressive from stable disease. The NETest was significantly lower in stable disease (34.1+2.6%) than in progressive disease (84+2.8%, p<0.0001). Integration of biologically relevant gene cluster expression differentiated stable from progressive disease. Combination of gene cluster data with the blood-algorithm (the NETest) predicted disease status in>92%. Blood transcript measurement predicts NET activity.

A multianalyte PCR blood test outperforms single analyte ELISAs for neuroendocrine tumor detection.

Modlin IM, Drozdov I, Alaimo D, Callahan S, Teixeira N, Bodei L, Kidd M.

A critical requirement in neuroendocrine tumor (NET) management is a blood biomarker test that is sensitive, specific and reproducible. We evaluated the NETest detect tumors, compared it with chromogranin A (CgA) and examined the confounding effect of proton pump inhibitors (PPIs), which cause falsely elevated CgA levels. NETest performance metrics were as follows: sensitivity 98.4%, specificity 100%, positive predictive value 100%, negative predictive value 97.8%, and the ROC-derived area under the curve (AUC) was 0.997. These were significantly better than CgA (all metrics <60%; AUC, 0.54; Z-statistic, 10.44, p<0.0001). The NETest was not elevated in any PPI-treated subject compared to 83% of CgA. The NETest is significantly more sensitive than plasma CgA for NET detection and is unaffected by acid suppression therapy.

A PCR blood test outperforms chromogranin A in carcinoid detection and is unaffected by PPIs

Modlin IM, Kidd M, Aslanian H, Bodei L, Drozdov I

A critical requirement in neuroendocrine tumor (NET) management is a sensitive, specific and reproducible blood biomarker test. We evaluated the NETest versus chromogranin A (CgA), pancreastatin (PST) and neurokinin A (NKA). NETest was >92% accurate. CgA was 76% accurate, PST 63% accurate and NKA 39% accurate. NETest significantly outperformed single analyte tests (area differences: 0.284–0.403, Z-statistic 4.85–5.9, P<0.0001). The NETest was the most sensitive and accurate tool for NET diagnosis. Its use will facilitate early detection of disease recurrence and can predict therapeutic efficacy.

The clinical utility of a novel blood-based multi-transcriptome assay for the diagnosis of neuroendocrine tumors of the gastrointestinal tract

Modlin IM, Kidd M, Bodei L, Drozdov I, Aslanian H

This study demonstrates that a blood-based multianalyte NET gene transcript measurement of well-differentiated small intestinal and pancreatic neuroendocrine tumor disease is sensitive (94-98%) and specific and outperforms the current monoanalyte diagnostic strategy of plasma CgA (52%) measurement.

The clinical utility of circulating neuroendocrine gene transcript analysis in well-differentiated paragangliomas and pheochromocytomas.

Pęczkowska M, Cwikla J, Kidd M, Lewczuk A, Kolasinska-Ćwikła A, Niec D, Michałowska I, Prejbisz A, Januszewicz A, Chiarelli J, Bodei L, Modlin I.

Paragangliomas and pheochromocytomas (PPGLs) exhibit variable malignancy, which is difficult to determine by histopathology, amine measurements or tissue genetic analyses. We evaluated the NETest as a diagnostic and prognostic for these tumors. Circulating NET transcript analysis is positive (100% diagnostic) in well-differentiated PCC/PGL, scores were elevated in progressive disease irrespective of mutation or biochemical activity and elevated levels were prognostic.

A liquid biopsy for bronchopulmonary/lung carcinoid diagnosis

Kidd M, Modlin IM, Drozdov I, Aslanian H, Bodei L, Matar S, Chung KM

No effective blood biomarker exists to detect and clinically manage bronchopulmonary (BP) neuroendocrine tumors (NET). We the blood-based 51 NET-specific transcript set for diagnosis and monitoring and evaluated clinical performance metrics in BPNETs. All 51 genes were identified in BPNET transcriptomes, tumor samples and cell-lines. Significant correlations were evident between paired tumor and blood (R2:0.63–0.91, p < 0.001). PCA and hierarchical clustering identified blood gene expression was significantly different between lung cancers and benign diseases, including BPNETs. Gene expression was highly correlated (R2: 0.91, p = 1.7 x 10-15) between small bowel and BPNET. For validation. Scores were significantly elevated (p < 0.0001) in BPNETs (57 ± 28%) compared to controls (4 ± 5%). BPNETs with progressive disease (85 ± 11%) exhibited higher scores than stable disease (32 ± 7%, p < 0.0001). Blood measurements accurately diagnosed bronchopulmonary carcinoids, distinguishing stable from progressive disease. This marker panel will have clinical utility as a diagnostic liquid biopsy able to define disease activity and progression in real-time.

The Utility of Blood Neuroendocrine Gene Transcript Measurement in the Diagnosis of Bronchopulmonary Neuroendocrine Tumours and as a tool to Evaluate Surgical Resection and Disease Progression

Filosso PL, Kidd M, Roffinella M, et al

The management of bronchopulmonary neuroendocrine tumors (BPNETs) is difficult, since imaging, histology and bio-markers have a limited value in diagnosis, predicting outcome and defining therapeutic efficacy. We evaluated a NET multi gene blood test (NETest) to diagnose BPNETs, assess disease status and evaluate surgical resection.

The results demonstrate that blood NET gene levels accurately identified BPNETs (100%) and differentiated these from controls, benign and malignant lung disease. Progressive disease could be identified and surgical resection verified. Chromogranin A had no clinical utility. Monitoring NET transcript levels in blood will facilitate management by detecting residual tumor and identifying progressive disease.

NET blood transcript analysis defines the crossing of the clinical Rubicon: when stable disease becomes progressive

Pavel M, Jann H, Prasad V, Drozdov I, Modlin IM, Kidd M.

A key issue in Gastroenteropancreatic neuroendocrine tumors (GEP-NETs) is early identification and prediction of disease progression. Clinical evaluation and imaging are limited due to the lack of sensitivity and disease indolence. We assessed the NETest as a predictive and prognostic marker of progression in a long-term follow-up (median 4 years) study. The NETest correlated with a well-differentiated GEP-NET clinical status. The NETest has predictive and prognostic utility for GEP-NETs identifying clinically actionable Alterations ~1 year before image-based evidence of progression.

The clinical utility of circulating neuroendocrine gene transcript analysis in well-differentiated paragangliomas and pheochromocytomas

Pęczkowska M, Cwikla J, Kidd M, Lewczuk A, Kolasinska-Ćwikła A, Niec D, Michałowska I, Prejbisz A, Januszewicz A, Chiarelli J, Bodei L, Modlin I

Spider plot identifying the changes from baseline. All three patients who continued to progress or developed progressive disease (arrows) exhibited a positive change in the NETest score. This who were stable or underwent curative surgery demonstrated a decrease in scores.
Arrows identify scores associated with clinically confirmed progressive disease.
Star identifies patient who underwent surgery.

Assessment of NETest Clinical utility in a US Registry-based study

Liu E, Paulson S, Gulati A, Freudman J, Grosh W, Kafer S, Wickremesinghe PC, Salem RR, Bodei L

Clinical relevance of molecular biomarkers in oncology management has been recognized in breast and lung cancers. The NETest was evaluate for management of neuroendocrine tumor disease (NETs) in a real-world study (USA registry (RegisterNET: NCT02270567). The NETest was the only feature linked to PFS (OR: 6.1, p<0.0001). High NETest correlated with progressive disease (96%; mPFS 6 months); low NETest with stable disease (94%; mPFS not reached). Use of NETest helped reduce imaging (biannual to annual) in 36-38%.

Circulating Transcript Analysis (NETest) in GEP-NETs Treated With Somatostatin Analogs Defines Therapy

Ćwikła JB, Bodei L, Cwikla A, Sankowski A, Alaimo D, Modlin IM, Kidd M.

Early and precise delineation of therapeutic responses are key issues in neuroendocrine neoplasm/tumor management. Imaging is currently used but exhibits limitations in sensitivity and specificity. The utility of biomarkers is unclear. We evaluated the NETest in somatostatin analog-treated GEP-NETs and compared it to CgA as a monitor of therapy. NETest values (>80%) were more accurate and occurred at a significantly earlier time point than CgA and predicted SSA treatment response.

Measurement of circulating transcripts and gene cluster analysis predicts and defines therapeutic efficacy of Peptide Receptor Radionuclide Therapy (PRRT) in neuroendocrine tumors

Bodei L, Kidd M, Modlin IM, Severi S, Drozdov I, Nicolini S, Kwekkeboom DJ, Krenning EP, Baum RP, Paganelli G

We evalutated the NETest prior to PRRT, during therapy and at 3 and 6 months after therapy in 54 177Lu-treated GEP-NET. The NETest accurately correlated with standard morphologic and functional imaging and therefore with treatment response and outcome of therapy.

NET blood transcript analysis defines the crossing of the clinical Rubicon: when stable disease becomes progressive

Pavel M, Jann H, Prasad V, Drozdov I, Modlin IM, Kidd M

Time prior to disease progression identified by imaging. A rise in NETest >70% occurred a median 1.62 years before imaging confirmation of tumor progression (failure of therapy). A rise in NETest >80% occurred a median 0.76 years before imaging confirmation of tumor progression (failure of therapy).

Assessment of NETest Clinical utility in a US Registry-based study

Liu E, Paulson S, Gulati A, Freudman J, Grosh W, Kafer S, Wickremesinghe PC, Salem RR, Bodei L

Relationship between NETest score and clinical management
A. In the watch-and-wait cohort, a low score was associated with in no treatment intervention in 93%. A high score led to a treatment intervention in 83%.
B. In the treatment cohort, a low score was associated with no change in treatment in 100%. A high score led to a treatment modification in 86%.
SD = stable disease PD = progressive disease

Blood measurements of Neuroendocrine Gene Transcripts Defines the Effectiveness of Surgical Resection and Ablation Strategies

Modlin IM, Frilling AF, Salem RR, Alaimo D, Drymousis P, Wasan HS, Callahan S, Faiz O, Weng L, Teixeira NS, Bodei L, Drozdov I, Kidd M

Surgery significantly reduced NETest levels consistent with removal of the source of the circulating gene expression.
In those with surgical cures (NED – no evidence of disease), elevated levels after surgery predicted disease recurrence.

The utility of blood neuroendocrine gene transcript measurement in the diagnosis of bronchopulmonary neuroendocrine tumours and as a tool to evaluate surgical resection and disease progression

Filosso PL, Kidd M, Roffinella M, Lewczuk A, Chung KM, Kolasinska-Cwikla A, Cwikla J, Lowczak A, Doboszynska A, Malczewska A, Catalano M, Zunino V, Boita M, Arvat E, Cristofori R, Guerrera F, Oliaro A, Tesselaar M, Buikhuisen W, Kos-Kudla B, Papotti M, Bodei L, Drozdov I, Modlin I

Current biomarkers used in BPNET management are single analytes and have a low utility, e.g., CgA (Chromogranin A) or NSE (neuron specific enolase). Histology and imaging have a high clinical utility. Imaging is especially useful in assessing disease evolution over time. Monoanalytes have minimal utility in assessing surgical residual disease or recurrence and limited benefit, except NSE in SCLC. Neuroendocrine circulating transcripts measurement (NETest – transcripts) is effective in all four stages of disease management.

Measurement of circulating transcripts and gene cluster analysis predicts and defines therapeutic efficacy of Peptide Receptor Radionuclide Therapy (PRRT) in neuroendocrine tumors

Bodei L, Kidd M, Modlin IM, Severi S, Drozdov I, Nicolini S, Kwekkeboom DJ, Krenning EP, Baum RP, Paganelli G.

We developed a complementary diagnostic based on circulating gene expression measurements (Ome index) and grading which can be used to predict response to PRRT. The accuracy of this Prediction Quotient in this discovery cohort of 54 NETs was 94%.

Therapy: The role of liquid biopsies to manage and predict PRRT for NETs

Kidd M, Modlin IM

Patients with neuroendocrine tumors are increasingly treated with peptide receptor radionuclide therapy. However, somatostatin receptor expression evaluation cannot predict who will respond to therapy. Additional criteria to identity which patients are most likely to respond and those who will develop radiation-associated sequelae are critical requirements. One mechanism are liquid biopsies – typically measurements of circulating RNA expression – which can be used as a complementary diagnostic for lutetium-based therapies.

PRRT genomic signature in blood for prediction of 177Lu-octreotate efficacy

Bodei L, Kidd MS, Singh A, van der Zwan WA, Severi S, Drozdov IA, Cwikla J, Baum RP, Kwekkeboom DJ, Paganelli G, Krenning EP, Modlin IM

We validated the PRRT Prediction Quotient (PPQ) as a complementary diagnostic in two different PRRT cohorts. The accuracy of this Prediction Quotient in the validation cohorts was 95%. It was significantly more predictive and of more clinical better than either grade (70%) or elevated chromogranin A (50%). Other parameters like SUVmax and FDG were equally non-informative.