Clinical Trial Details
— Status: Recruiting
Administrative data
| NCT number |
NCT05399719 |
| Other study ID # |
202201057RINA |
| Secondary ID |
|
| Status |
Recruiting |
| Phase |
|
| First received |
|
| Last updated |
|
| Start date |
May 25, 2022 |
| Est. completion date |
March 10, 2025 |
Study information
| Verified date |
June 2023 |
| Source |
National Taiwan University Hospital |
| Contact |
Hui Ju Ch'ang, MD |
| Phone |
886-0911200099 |
| Email |
hjmc[@]nhri.org.tw |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
Clinical and preclinical evidence reveal that cancer cells may fuse with hematopoietic cells
to obtain properties including migration, proliferation and drug resistance. The
investigators hypothesize that cancer cell-macrophage fusion hybrids may lead to pancreatic
cancer desmoplasia and progression.
Murine tumor models using cre-loxP or gender-mismatched xenografts as well as
pdx-cre-KrasLSL-G12D mice after bone marrow transplantation from reporter ROSA mice were
established. Fusion hybrids and macrophage markers were detected using immunofluorescence
staining and flowcytometry.
In vitro co-culture using cre-loxP or dual fluorescence methods of pancreatic cancer cells
with macrophages was used to evaluate the frequency of fusion phenomenon. The proliferative,
migratory and resistant phenotypes of purified fusion hybrids were measured. Differentially
expressed genes between fusion hybrids and non-fused cancer cells were compared by Affymetrix
microarray analysis.
The investigators are going to collect tumor tissues from cancer patients who received
allographic bone marrow transplantation before. We will evaluate Y chromosome or short tandem
repeats to identify donor- derived genes in cancer cells and demonstrate the clinical
evidence of fusion between cancer cells and macrophages. The tumor tissues will be collected
from the Pathology Department. Ten slides of 4-8um will be collected from twenty patients
enrolled according to the inclusion criteria.
The investigator will collect peripheral mononuclear cells from healthy volunteer ( eg.
Donors for bone marrow transplantation) or hyperemia patients. The mononuclear cells will be
induced to differentiate into macrophages and will be co-cultured with cancer cells in order
to purify fusion hybrids. The fusion hybrids between cancer cells and macrophages will be
evaluated for biologic characters including proliferation, radio-sensitivity, migration etc.
The investigators planned to collect blood samples from Department of Laboratory Medicine,
Blood bank. Thirty subjects of healthy volunteer or hyperemia patients will be enrolled. Ten
to 20ml peripheral blood will be collected from each subjects for one time.
Description:
Desmoplasia has been shown to exert mechanical forces and create a biochemical intra-tumoral
immunity and influence the development and progression of a malignancy. Recent reports
suggest a significant negative correlation between cancer patient survival and extracellular
matrix deposition in primary tumors; and concomitant stromal targeting may enhance
therapeutic outcomes in cancer patients.
In defining the source of the dense stroma, it is generally believed that over-activated CAFs
could produce extracellular matrix (ECM) proteins, the major component of the dense stroma.
Anti-fibrosis drugs including those depleting CAFs or collagen, sonic hedgehog inhibitor,
antibodies against LOX/LOX2, were under clinical trials. However, CAF-depleted tumors
displayed a more aggressive phenotype and alteration in regulatory T cells, raising the
possibility that CAF-targeting strategies may have both beneficial and detrimental effects.
Considering the functional significance of macrophages in inflammation, desmoplasia, cancer
initiation and progression, mounting studies had conducted to evaluate the efficiency of
anti-macrophage as a novel strategy against cancer. Accordingly, the anti-macrophage
strategies include the inhibition of the monocytes recruitment as well as transformation, and
the ablation the macrophages directly. Given the multifaceted roles of maintaining
homeostasis, the systemic depletion of macrophages may lead to increased infections or
impaired ability of tissue-resident cells to carry out their normal function. Thus, the
identification of tumor associated macrophage (TAM)-specific markers will enable the
development of more sophisticated therapies that can be targeted specifically to tumors
without affecting the function of other tissue-resident immune cells.
Recent studies had identified numerous factors such as platelet-derived growth factor (PDGF)
and transforming growth factor b1 (TGFb1) released by TAMs, especially bone marrow
(BM)-derived ones, involving in the process of activating CAFs. In addition, there were
TAMs-derived pro-fibrotic factors, such as connective tissue growth factor (CTGF), chemokine
ligand 17 (CCL17), CCL22 and reactive oxygen species (ROS). Similar observation was found in
TAMs-derived matrix metallopeptidase 9 (MMP9) which involved in the stroma turnover by
degrading the ECM proteins. However, rare study investigates the role of fusion between
macrophages and cancer cells in desmoplasia and tumor progression as well as the strategy
targeting fusion machinery of macrophages with cancer cells.
Several lines of evidence support that cell fusion between cancer cells and leukocytes,
majorly macrophages, is one of the explanations for tumor resistance and progression. Recent
genotyping of a metastatic melanoma to the brain that arose following allogeneic BM
transplantation supports, first time clinically, fusion between a BM derived cell and a tumor
cell playing a role in the origin of metastasis. Fusion hybrids enumerated in peripheral
blood of pancreatic cancer patients were reported to correlate with disease stage and predict
overall survival. Furthermore, general inflammatory responses were demonstrated to increase
the number of fusion events. While the fusion events were demonstrated in animal and human,
and were correlated with cancer resistance and metastasis, little is known regarding key
questions such as the mechanisms through which macrophage-cancer cell fusion and subsequent
genomic hybridization occurs in vivo; potential survival advantages of hybrids; reprogramming
of fibrotic and immunologic tumor microenvironment by hybrids; and the development of novel
strategies to target fusion machinery.
The majority of macrophages, including tissue macrophages in normal tissue and TAMs, are
derived from bone marrow (BM). The investigators' preliminary study, using irradiated murine
intestine model, suggested parallel between the development of cell fusion between BM-derived
monocytes and intestine stromal cells with radiation-induced fibrosis. Using cre-loxP system
and gender mismatched BM transplantation, we demonstrated that BM-derived CD11b(+)
myelo-monocytic cells/macrophages were the major fusion partner to damaged intestine stromal
cells after radiation. The fusion events in irradiated intestine stroma correlated with
chronic intestine fibrosis with enhanced TGFb transcripts and collagen deposition. With
macrophage deletion by liposomal clodronate or conditional CD11b knock-out, we can ameliorate
radiation induced chronic intestine fibrosis.
The main property of cancer cells that makes them malignant is the ability to produce diverse
progeny. Fusion between cancer cells and TAMs can be an engine of genomic and epigenetic
variability that has a potential to make cells with new properties at a rate exceeding that
achievable by random mutation. Previous reports revealed cell fusion contribute to drug
resistance and can promote the ability of cancer cells to metastasize.
Fibrosis and inflammation are hallmarks of tumor desmoplasia. TAMs, majorly derived from bone
marrow,5 act as an essential connecting moiety between inflammation and cancer via secretion
of pro-inflammatory cytokines/chemokines. The M2 polarized macrophages, which constitute
majority of the macrophages in tumor microenvironment, secrete IL10 and other cytokines that
mediate T helper 2 (Th-2) responses and are responsible for malignant tumor transformation
and inhibit antitumor immune response mediated by T cells. TAMs were reported to play a
phenomenal role in enhancing mesenchymal phenotype of cancer cells via TGFb. The
lipopolysaccharide (LPS) receptor toll-like receptor 4 (TLR4) on the surface of TAMs has been
implicated in a role in the EMT via TLR4/Interleukin 10 (IL10) cascade. The level of secreted
protein acidic and cysteine-rich (SPARC) gene, a regulator of EMT, was reported to be 3-4
fold higher in hybrids of BM-derived cell and melanoma.
Analysis of microenvironment microarray assay revealed that fusion hybrids of colon cancer
cells and macrophages exhibited a combination of biases, reflecting properties of both
parental cells, including fibronectin, collagen XXIII, vitronectin, potentially providing a
broader desmoplastic process in different microenvironment. The investigators' preliminary
results of differential expression of genes between fusion hybrids of cancer cells and
macrophages with non-fused cancer cells show significant elevation of signal pathways
involving inflammation, extracellular matrix degradation and mesenchymal phenotypes etc.
Besides, prominent increase in signal regulatory protein (SIRP) family members, a cell-cell
communicating system, were also found.
Current treatment result of cancer with desmoplasia is unsatisfactory. It is generally
believed that desmoplastic stroma provides cancer cells with cytokines/chemokines to suppress
immune surveillance and to enhance malignant progression. Recent clinical trials revealed
direct targeting desmoplasia using sonic hedgehog inhibitor lead to a more aggressive
phenotype with increased regulatory T cells. On the other hand, depleting macrophage or its
recruitment from circulation revealed to be effective in reducing pro-fibrotic
cytokines/chemokines and CAFs activity; however, with significant normal tissue toxicity.
From previous investigation and our study, fusion hybrids between cancer cells and
macrophages appeared to play important role in promoting cancer desmoplasia as well as
malignant progression. The investigator proposed to develop an alternative method to
ameliorate cancer desmoplasia by focusing on novel targets interfering fusion between cancer
cells and macrophages. The potential candidates were revealed by our preliminary microarray
study. eg. Signal regulatory protein (SIRP) signal pathway including SIRPa, a macrophage
fusion receptor, which was nearly 5 folds differentially expressed in fusion hybrids.
Furthermore, the down-stream molecules including DAP12 (DNAX activating protein), a general
macrophage fusion regulator, and protein tyrosine phosphatase (PTPN6), protein tyrosine
kinase (PTK2B) were increased 8 folds and 4 folds, respectively, in fusion hybrids.
We proposed to target on SIRPa, and other potential candidate molecules with neutralizing
antibodies or silencing mRNA to reduce the incidence of fusion phenomenon in cancer with
desmoplasia. By reducing the frequency of tumor heterogeneity and the fibrotic phenotype, we
would like to improve the therapeutic outcome of cancer with desmoplasia. The combination
effect of anti-fusion agents with other anti-fibrosis agents or immunologic therapy will also
be explored
