The Securities and Exchange Commission has not necessarily reviewed the information in this filing and has not determined if it is accurate and complete.
The reader should not assume that the information is accurate and complete.

UNITED STATES SECURITIES AND EXCHANGE COMMISSION
Washington, D.C. 20549
FORM 13F

FORM 13F COVER PAGE

OMB APPROVAL
OMB Number: 3235-0006
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Report for the Calendar Year or Quarter Ended: 09-30-2021
Check here if Amendment    Amendment Number:
This Amendment (Check only one.):    is a restatement.
   adds new holdings entries.
Institutional Investment Manager Filing this Report:
Name: TWO SIGMA INVESTMENTS, LP
Address: 100 Avenue of the Americas
16th Floor
New York, NY  10013
Form 13F File Number: 028-10385

The institutional investment manager filing this report and the person by whom it is signed hereby represent that the person signing the report is authorized to submit it, that all information contained herein is true, correct and complete, and that it is understood that all required items, statements, schedules, lists, and tables, are considered integral parts of this form.

Person Signing this Report on Behalf of Reporting Manager:
Name: Matthew B. Siano, Esq.
Title: General Counsel
Phone: 212-625-5700
Signature, Place, and Date of Signing:
/s/ Matthew B. Siano, Esq. New YorkNY 11-15-2021
[Signature] [City, State] [Date]
This Form 13F holdings report is being filed on behalf of Two Sigma Investments, LP ("Investments"). Investments serves as the sole manager or investment manager, as applicable, of (i) Two Sigma Equity Portfolio, LLC ("Equity"), Two Sigma Spectrum Portfolio, LLC ("Spectrum"), Two Sigma Futures Portfolio, LLC ("Futures"), Two Sigma Horizon Portfolio, LLC ("Horizon"), Two Sigma Options Portfolio, LLC ("Options") and Two Sigma Equity Spectrum Portfolio, LLC ("Equity Spectrum"), six investment entities that are managed by Investments, each of which may be deemed to be an "other manager" for purposes of this Form 13F report and (ii) Two Sigma Galileo Portfolio, LLC ("Galileo"), an investment entity that is managed by Investments and that may not presently be deemed to be an "other manager" for purposes of this Form 13F report. Investments, rather than Equity, Spectrum, Futures, Horizon, Options, Equity Spectrum or Galileo, exercises sole investment discretion and voting authority over the securities held in the various accounts of Equity, Spectrum, Futures, Horizon, Options, Equity Spectrum and Galileo.
Report Type (Check only one.):
X 13F HOLDINGS REPORT. (Check here if all holdings of this reporting manager are reported in this report.)
   13F NOTICE. (Check here if no holdings reported are in this report, and all holdings are reported by other reporting manager(s).)
   13F COMBINATION REPORT. (Check here if a portion of the holdings for this reporting manager are reported in this report and a portion are reported by other reporting manager(s).)
Form 13F Summary Page
Report Summary:
Number of Other Included Managers: 6
Form 13F Information Table Entry Total: 4,175
Form 13F Information Table Value Total: 44,745,686
(thousands)
List of Other Included Managers:
Provide a numbered list of the name(s) and Form 13F file number(s) of all institutional investment managers with respect to which this report is filed, other than the manager filing this report.
[If there are no entries in this list, state “NONE” and omit the column headings and list entries.]
No. Form 13F File Number Name
1 028-10704 TWO SIGMA EQUITY PORTFOLIO LLC
2 028-11144 TWO SIGMA SPECTRUM PORTFOLIO, LLC
3 028-16011 Two Sigma Futures Portfolio, LLC
4 028-15356 TWO SIGMA HORIZON PORTFOLIO, LLC
5 028-17336 Two Sigma Options Portfolio, LLC
6 028-20138 Two Sigma Equity Spectrum Portfolio, LLC