Clinical Study of Granulocyte Infusion for Advanced Cancer

Background \& Rationale:

For years, most tumor immunotherapy researches have focused on T cell and natural killer (NK) cell therapies, most of which involve amplification and modification of the patient's immune cells for reinfusion therapy. However, for the treatment of solid tumors, there is currently little breakthrough. Recently, researchers have reported a colony of cancer-resistant mice developed from a single mouse that was immune to multiple lethal cancer cell injections. Further research revealed that such anti-cancer immunity can cause rapid shrinkage or disappearance of the tumors in other cancer-bearing mice. Interestingly, this therapeutic effect is due to the donor granulocytes, instead of T cells or NK cells. Infusion of granulocytes is a classic therapy in treating infection associated with granulocytopenia. Currently, clinical collection of blood components, including isolation of granulocytes, is a mature technique. The infusion of granulocytes is a viable anticancer therapy combining the classic technique and novel anticancer approach. This proposed trial will test whether granulocyte infusions from healthy unrelated donors can be used to treat advanced cancer. In the proposed trial, up to 100 Subjects with advanced cancer can be entered. Each patient will be given a dose of (2.0-5.0)x10\^10 granulocytes from a different healthy donor every week over a course of 5 doses. The trial will evaluate the subject's cancer 7, 30, 90 and 180 days after the last infusion.

Granulocyte anti-cancer therapy refers to a method in which healthy donor granulocytes with high cancer-killing activity are collected and infused into a specific cancer patient by matching, to achieve the therapeutic anticancer effect. In this proposed trial, up to 100 Subjects with advanced cancer can be entered. Potentially hundreds of healthy Donor-participants will be recruited. First, granulocyte donors will be identified via in-vitro assay of Cancer Killing Activity (CKA), which screens healthy Donor-participants for those with anticancer activity of more than 80%. Secondly, after donor-recipient blood matching, (2.0-5.0)x10\^10 granulocytes will be collected from each donor, accounting for about 1/10 of the human body. Each patient will be given a dose of (2.0-5.0)x10\^10 granulocytes from a different donor per week over a course of 5 doses (with an ideal total infusion of 2X10\^11 granulocytes). After each infusion, the patients will be monitored carefully for possible adverse events. If adverse events occur, the infusion can be slowed down or stopped until the adverse events can be managed. The trial will observe the subject's cancer 7, 30, 90 and 180 days after the last infusion. Target lesions, non-target lesions, and new lesions will be evaluated via medical imaging and tumor markers. The responses will be compared against the measurements at baseline.

Eligibility Criteria:
Inclusion Criteria for Subjects:

* Malignant tumor confirmed by puncture pathology/postoperative pathology;
* Clinical stage IV;
* Performance status of ≤2 on the ECOG scale
* Life expectancy of at least 3 months
* All patients was diagnosed metastatic tumor by histology and cytology. Cytoreductive surgery can be performed if the patient can endure surgery.
* Measurable Disease: Lesions that can be accurately measured in at least one dimension (longest diameter recorded) as 20 mm with conventional technique or as 10 mm with spiral CT scan.If the lesion is between 15mm and 20mm, the thickness of the CT layer should be no more than 0.5mm. For multiple lesions, select a representative 10 lesions (maximum 5 in the same organ) and calculate the sum of the longest diameters of all target lesions as the baseline sum diameter.
* ≥ 4 weeks since prior medical therapy, radiation therapy, and surgery
* Laboratory tests meet the following criteria:

A. Bone marrow function: Absolute blood neutrophil (ANC) count ≥1\*10\^9 /L, blood small (PLT) ≥75\*10\^9 /L.

B. Liver function: serum total bilirubin (STB), combined bilirubin (CB) ≤ upper limit of normal (ULN) \* 1.5, alanine aminotransferase (ALT), aspartate aminotransferase (AST) ≤ULN\*2.5 (in the absence of liver metastases), or ≤ULN\*5 (with liver metastases);

C. Renal function: serum creatinine (Cr) ≤ ULN \* 1.5, endogenous creatinine clearance (Ccr) ≥ 50 mL / min (calculated using the Cockcroft-Gault formula, see Appendix 2);

* The anti-neutrophil antibody test result was negative.
* The patient volunteered and signed an informed consent form.

Exclusion Criteria of Subjects:

* Arrhythmia, congestive heart failure, or severe coronary artery disease;
* Pregnant or nursing women;
* Patients with severe autoimmune diseases;
* Patients who have been using or are using immunosuppressive agents for a long time;
* Those with coagulopathy;
* Cases that the investigator considered inappropriate for inclusion.

Inclusion Criteria for Granulocyte Donors:

* Must have signed Donor-participant Informed Consent Form
* Must be able to provide granulocytes in local blood donation center, and transport collected granulocytes to the hospital blood bank or clinical application within 24 hours;
* Must score \>80% in Cancer Killing Assay (CKA), a score of 60-80% would be considered when there are insufficient donors;
* Must have CMV negative or positive sero-testing completed; only seronegative donors are accepted for a seronegative recipient;
* Must have compatible ABO and RH typing with the recipient;
* Must be qualified for blood donation: aged 18-35, male \>50kg or female \>45kg, normal cardiopulmonary function, specific gravity of blood male≥1.052 or female≥1.051, HbsAg, anti-HCV, anti-HIV, or syphilis tests.

Exclusion Criteria for Granulocyte Donors:

* Dental or other minor surgery within preceding 15 days;
* Arrhythmia, congestive heart failure, or severe coronary artery disease;
* Major surgery within preceding 6 months;
* Appendectomy, hernia repair, or tonsillectomy within preceding 3 months;
* Females 3 days before or after menstruation, ≤6 months since prior pregnancy or abortion, ≤ 1 year since delivery/breastfeeding;
* Recovery from flu or acute gastroenteritis within preceding week, acute renal infection within preceding month, pneumonia within preceding 3 months;
* Recovery from infectious diseases such as dysentery within preceding 6 months, typhoid fever within preceding year, brucellosis within preceding 2 years, or malaria within preceding 3 years;
* Recovery from localized dermatitis within preceding week, extensive inflammation within preceding 2 weeks;
* Receiving whole blood or blood components within preceding 5 years;
* Allergic to G-CSF mobilizer.