Evolution of Metabolic and Immune Dysfunction in In-transit Melanoma
ConditionMelanoma
Trial statusRecruiting
Trial phaseNot listed
Trial typeObservational
Biological sexAll
Age18+
SponsorYana Najjar
About this trial
Melanoma in-transit metastases (ITMs) continue to represent a therapeutic dilemma, in that no standard method of treatment has been uniformly adopted. The complexity and heterogeneity of patient and disease characteristics, including the location and number of ITMs presents a barrier to a one size fits all treatment approach. Treatment of patients with limited regional disease remains challenging. Patients are typically treated with a combination of surgery, regional therapy, systemic therapy. Data on the management of ITMs is limited, even with the availability of immunotherapy (IMT). This study will use the unique etiology of ITMs to facilitate the understanding of how individual lesions metabolically and immunologically evolve as they move away from the primary tumor site. It is hypothesize that as ITMs move away from the primary melanoma site each will harbor progressively hypermetabolic tumor cells and a harsher microenvironment.
Eligibility criteria
This trial does not accept healthy volunteersQualifiers
Be willing and able to provide written informed consent for the trial.
Be ≥ 18 years of age on day of signing informed consent.
A histological diagnosis of melanoma and at least two in-transit lesions at distinct distances from the primary site. Patients may be enrolled on the basis of a diagnosis of in-transit disease by a treating melanoma oncologist.
Cutaneous, mucosal or uveal melanoma are permitted.
Disqualifiers
Subjects with in-transit disease that is not amenable to biopsy per the treating physician are excluded.
Subjects with known chronic immunosuppression (such as biologic agents like remicade, mycophenolate, methotrexate, prednisone >20 mg daily).
Subjects who are known to be HIV+, Hep B or Hep C positive.
Trial population
Patients with a histological diagnosis of melanoma
Trial design
Design model
Cohort
Time perspective
Prospective
Treatments tested in this trial
Pimonidazole
DrugPimonidazole is not used with therapeutic intent, and has a non-hazardous designation. It has been widely used for in-vivo evaluation of intratumor hypoxia, and patients will take PO pimonidazole before the scheduled biopsy. Patients receive an oral dose of pimonidazole, a safe chemical tracer up to 24 hours prior to biopsy. Pimonidazole allows for true hypoxia staining; pimonidazole binds hypoxic proteins covalently, creating an antigen that facilitates the imaging, flow cytometry, and scRNA-seq experiments proposed. Pimonidazole has been previously used in patients and is safe and well tolerated, without anticipated adverse events.
Treatment groups
20 Participants
are divided into 1 treatment groupGroup A: Pimonidazole1 intervention
Trial outcomes
Primary outcomes
1
Immunometabolic profiling
Immunometabolic profiling of individual microenvironments in ITMs (in-transit metastases) using flow cytometry via T cell subsets, markers of activation/exhaustion, and metabolic insufficiency (mitochondrial mass, glucose uptake capacity, and hypoxia by pimonidazole staining.
Time frame
At baseline2
Tumor cell metabolism
Tumor cells metabolism of ITMs (in-transit metastases) will be profiled using the Seahorse flux analyzer to measure oxygen consumption rate (OCR) and extracellular acidification rate (ECAR).
Time frame
At baseline3
Imaging of individual ITM stations
CODEX imaging will be conducted on sections from each station using extensive phenotyping panels.
Time frame
At baseline4
hypoxia exposure analyses
Akoya Biosciences' CODEX analysis suite and custom ImageJ plugins will be used to determine proximity of cells to one another, coincidence with hypoxic areas, and accumulation of regulatory/dysfunctional populations.
Time frame
At baselineSecondary outcomes
1
Whole exome sequencing of tumor cells
Whole exome sequencing will be performed to identify mutations in tumor cells isolated from each lesion. Analysis will be in an 'intrapatient' manner as a function of distance from the primary site.
Time frame
At baseline2
Clonal evolution analysis of individual ITMs
Mutations will be used to construct trees revealing clonal representation and diversity at each station. The goal is to determine how migration away from the primary site affects a tumor's heterogeneity in terms of tumor cells.
Time frame
At baseline3
Cellular heterogeneity and transcriptomic state
The composition of each ITM tumor microenvironment will be analyzed using dimension reduction strategies, in collaboration with a bioinformatics core. Each biopsy sample will be sequenced simultaneously but hashed separately, allowing all ITMs to be compared in one run. Clusters will be identified using lineage-defining genes and then transcriptomic states identified within each cluster.
Time frame
At baseline4
TCR clonotype and trajectory analysis
5' scRNAseq will be used for identification of TCR sequences to identify individual T cell clones. Clonotypes will be revealed in each lesion, and diffusion pseudotime analyses will be employed within each ITM and between ITMs to determine differentiation states of various T cell subsets. It is thought that distant lesions will harbor more terminally differentiated T cells.
Time frame
At baselineSponsors and contacts
Click on the lead sponsor to view all of their trials.
Source ClinicalTrials.gov